An Introduction toBiotechnology

Amgen is a leading human therapeutics company in the biotechnology industry. Since our founding in

1980, we have focused on accomplishing our mission to serve patients by discovering, developing

and delivering innovative medicines to treat grievous illnesses. By pioneering the development of

novel products based on advances in cellular and molecular biology, Amgen’s therapeutics have

changed the practice of medicine and helped millions of people around the world to fight cancer,

kidney disease, rheumatoid arthritis and other serious illnesses.

Pioneering science delivers vital medicines

Chapter One: Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

Chapter Two: The Science . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

Chapter Three: How Biology Drives Biotechnology . . . . . . . . . . 9

Chapter Four: The Technology . . . . . . . . . . . . . . . . . . . . . . . . . . 13

Chapter Five: Drug Discovery . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

Chapter Six: Drug Development . . . . . . . . . . . . . . . . . . . . . . . . . 22

Chapter Seven: Scale-Up and Manufacturing . . . . . . . . . . . . . . 26

Chapter Eight: Biotechnology Medicines . . . . . . . . . . . . . . . . . . 29

Chapter Nine: Future of Biotechnology in Healthcare . . . . . . . . 31

Glossary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36

Timeline of Medical Biotechnology . . . . . . . . . . . . . . . . . . . . . . 42

Table of Contents

In 1919, Karl Ereky, a Hungarian engineer, coined the term biotechnology* to describe the

interaction of biology and human technology. He envisioned a new era of technology based on

using biology to turn raw materials into socially useful products. Nearly a century later, Ereky’s

vision is being realized by thousands of companies and research institutions.

Chapter One:

Introduction

Introduction *Terms in boldface are defined in the glossary .2

Introduction

Modern biotechnology began in the 1970s

in Northern California and has since grown

into a worldwide industry . Amgen, founded

in 1980, was one of the first companies to

realize the new field’s promise by bringing

biotechnology medicines to patients .

Today, biotechnology industry sectors include

healthcare (biologics, devices, diagnostics),

agriculture (genetically modified organisms,

food safety), industry and environment

(biofuels, biomaterials, pollution) and biode-

fense (vaccines, biosensors). This booklet

focuses on healthcare .

Drugs

A drug is a therapeutic substance used to

prevent, manage or cure disease . In the

United States, the U .S . Food and Drug

Administration (FDA) must approve all drugs

before they are sold to the public . Most

countries follow global harmonized guide-

lines and have a regulatory agency similar

to the FDA that evaluates drug research

and approves drugs for marketing . The most

familiar type of drug is the synthesized drug,

such as aspirin . The pharmaceutical industry

traditionally manufactures synthesized drugs .

The biotechnology revolution brought about

a new class of drug: the biologic .

Biologics are therapies derived from living

organisms and include therapeutic proteins,

DNA vaccines, monoclonal antibodies and

peptibodies [a modality that combines the

active portion of a protein (peptide) with a

portion of the core structure of an antibody],

as well as new experimental modalities such

as gene therapy, stem cell therapy, antisense

nucleotides and RNA viruses .

Many biotechnology drugs are proteins.

Proteins, which are made from amino acids,

are the workhorses of the cell and perform

all functions within a cell . Because cells

produce proteins naturally, the biotechnology

industry utilizes cells, not chemicals, to

manufacture biologics .

BIoFaCT

To bring a new drug to market (from discovery through clinical trials and FDA approval) costs an estimated $1 billion and can take 10 to 15 years or longer .* Only one in 10 new drugs that makes it into human testing actually makes it to market . Given this high failure rate and the tremendous cost of bringing a new therapy to market, companies depend on successful drugs to produce enough revenue to compensate for both the R&D costs of the successful therapies and the expense of failed ones .

* Innovation .org . (February 2007) . Drug discovery and development: Understanding the R&D process [Brochure] . Washington, DC: Pharmaceutical Research and Manufacturers of America . 3

The Science

The biotechnology industry is based on living organisms. The cell is the basic unit of life. All living

organisms consist of one or more cells. Some organisms are unicellular, such as bacteria and

yeast. Others, such as humans, are multicellular, consisting of trillions of cells. All cells have

common processes they perform in order to survive. Biotechnology harnesses these processes

to make products to treat illness and improve health.

Chapter Two:

The Science

4

The Science

Cell Processes

Cells Replicate (mitosis): Prior to dividing,

a cell makes a copy of its DNA and other cell

parts . The cell divides and forms two identical

cells from the original single cell . These iden-

tical cells are referred to as daughter cells .

Cells Grow: After replicating, the daughter

cells grow to their intended size .

Cells Metabolize: Cell metabolism is the

process by which cells process nutrients and

maintain a living state . Cells break down large

molecules into smaller molecules to produce

energy and molecular building blocks, which

are used to create new cell structures and

control cell function .

Cells Respond to Stimuli: Unicellular and

multicellular organisms respond to internal

and external stimuli . For example, plants grow

toward a light source because light is needed

for photosynthesis and the production of

energy . Light and the ability to respond to its

presence are essential to a plant’s survival .

Cells can respond to a whole range of stimuli .

Cells Adapt: Organisms may thrive or die

based on their ability to adapt to adverse

environmental conditions such as changes

in temperature, solute concentration, oxygen

supply and the presence of hazardous agents .

Parts of an Animal Cell

The cell can be divided into three main sections:

the cell membrane, the cytoplasm (which

includes the organelles) and the nucleus.

Nucleus: The nucleus houses most of the

DNA and is the control center of the cell . It is

surrounded by a membrane that lets certain

molecules into and out of the nucleus to

keep the DNA safe . This DNA never leaves

the nucleus .

Cell Membrane: The cell membrane is the

border that surrounds a cell . It monitors what

goes into and out of the cell . Embedded in the

cell membrane are receptors. Receptors

cross the membranes and act as docking

stations for molecules . When a specific type

of molecule attaches to a receptor, a series

of chemical reactions (cell signaling) can occur

in the cell, creating a cellular response . If a

receptor is blocked, cell signaling is stopped

and no response occurs . This is how some

biologic therapies work: they attach to recep-

tors and interrupt cell signaling . Other biologics

work by mimicking the signaling molecule .

Organelles: There are many different types

of organelles within the cytoplasm of a cell, and

each performs a specific function . For exam-

ple, ribosomes make proteins . Mitochondria

make energy . The endoplasmic reticulum folds

and transports certain proteins . Golgi bodies

modify proteins and are involved in their

transportation around the cell . Vacuoles

store cellular waste products for disposal .

BIoFaCT

Ribosomes can assemble an average-size protein in one minute .

NucleusNucleusCell Membrane

Cell MembraneCytoplasm

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DNA

All cells contain deoxyribonucleic acid (DNA).

DNA is the blueprint for the construction

and operation of the cell . DNA is arranged

in large segments called chromosomes.

Within a chromosome are specific pieces

of DNA called genes. Genes vary in length

and contain the information a cell needs to

make proteins . Proteins control many aspects

of cellular function .

The information in DNA is stored as a code

made up of four basic “building blocks”

called nucleotides . DNA is very long, and

the order of the nucleotides determines the

information stored . Each nucleotide consists

of three components: a deoxyribose sugar, a

phosphate group and a base . There are four

different types of bases: adenine (A), thymine

(T), guanine (G) and cytosine (C) . The order of

the As, Ts, Gs and Cs in DNA gives meaning

to the cell, just as the order of letters in a word

gives meaning to that word and, by extension,

to a story in which the word appears . The

diversity of organisms is a result of the limit-

less combinations of bases—As, Ts, Gs and

Cs . Every organism contains DNA, but the

number and arrangement of bases are

different for every organism .

DNA is called a double helix because it is

posited to consist of two strands of nucle-

otides that bond together in a very specific

manner . The As bond with Ts, and the Cs

bond with Gs . The resulting A-T and C-G

combinations are called base pairs. In each

human cell, the length of DNA is equal to 3

billion base pairs . If flattened, a DNA segment

would look something like a ladder with two

side rails (phosphate and ribose groups)

and rungs (base pairs) between them . The

structure makes DNA very stable and able

to carry vast amounts of information .

Most cells within an organism contain the

exact same DNA, but not all genes within each

cell are active, or turned on . When a gene

gets turned on, the information encoded by

the gene is used to produce, or express, the

Determining the Structure of DNA In 1962, James Watson, Francis

Crick and Maurice Wilkins jointly

received the Nobel Prize in Physi-

ology or Medicine for discovering

the structure of deoxyribonucleic

acid (DNA). Because the Nobel

Prize is awarded only to the living,

Wilkins’s colleague Rosalind

Franklin, who died from cancer

at the age of 37, could not be

honored. But many attribute the

success of Watson and Crick’s

1953 discovery to Franklin, whose

X-ray crystallography images

of DNA helped them clarify the

structure of DNA.

BIoFaCT

Mitochondria are the only organelles in ani-mal cells to have their own DNA . Mutations in mitochondrial DNA can lead to illnesses such as Kearns-Sayre syndrome, which causes the loss of heart, eye and muscle movement functions .

The molecular structure of DNA—the double helix.

BIoFaCT

Humans have 23 pairs of chromosomes, for a total of 46—half inherited from the mother and half from the father . Chromo-somal abnormalities can involve numeric abnormalities, where there may be more or less than the normal 46 chromosomes, or structural abnormalities, where there may be duplications, translocations, deletions or even inversions of sections of certain chromosomes .

The Science

ChromosomeGene

DNA

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encoded protein . When a gene is not active,

or is turned off, it is not used to express

proteins . Depending on the cell’s function and

needs, genes are either turned on or off . Many

diseases are the results of genes improperly

turned on or off .

Mutations

Any change in the DNA sequence is called

a mutation . Environmental factors such as

exposure to radiation or chemical toxins can

cause mutations .

Mutations also can occur during the natural

process of DNA replication, when the cell is

responsible for copying 3 billion base pairs in

20 hours . Bases can be substituted, deleted

or repeated . Changes in the DNA sequence

may cause proteins to become dysfunctional

or may even, occasionally, improve function .

Genetic diversity results from an accumulation

of mutations over a long period of time, which

causes the differences among species .

Genomes

The term genome refers to the entire genetic

information in an organism . The human genome

is the entire DNA content found in a human;

the corn genome is the entire DNA content

found in corn; and so on . All genomes are

made up of the same bases: As, Ts, Gs and

Cs . The differences between genomes lie in

the number and sequence of base pairs and

the number and sequence of genes .

The number of base pairs does not correspond

to the number of genes; the two are indepen-

dent of each other . For example, the human

genome has 3 billion base pairs and approxi-

mately 20,000 to 25,000 genes . Only 3 percent

of the human genome codes for genes; 97

percent is termed noncoding DNA—in other

words, DNA that does not contain instructions

for creating proteins . Biologists have not yet

fully discovered the function of noncoding

DNA, but they suspect it may be involved in

the evolution of species or may have regulatory

functions within the cell .

Furthermore, the number of base pairs and

genes does not correspond to the intelligence

or physical capability of an organism . Compare

the human genome with the amoeba genome .

An amoeba, a unicellular organism, has the

largest-known genome—which may be as

large as 670 billion base pairs—as compared

to the human genome of 3 billion base pairs .

Yet a human is a more complex, intelligent

and physically capable organism .

Proteins

Proteins are made of long chains of amino

acids that fold into intricate and complex

3-D shapes . The order of amino acids is

determined by the DNA sequence in a gene .

There are 20 different amino acids . The se-

quence of amino acids determines the shape,

and therefore the function, of the protein .

The U.S. National Center for Biotechnology Information Established in 1988, the National

Center for Biotechnology Informa-

tion (NCBI) creates and maintains

extensive networks of biomedical

databases for storing and analyzing

genomes. The databases include

information about various genome

base pair sequences, genes and

proteins.

NCBI is a division of the U.S.

National Library of Medicine at

the National Institutes of Health.

See http://www.ncbi.nlm.nih.gov.

BIoFaCT

Variations in individual nucleotides occur within DNA at the rate of approximately one in every 1,300 base pairs in most organisms . In humans, the rate is one in every 1,200 base pairs . Most of these mutations, how-ever, do not adversely affect us; only a few are involved in the production of dysfunc-tional proteins or disease states .

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The Science 7

Protein synthesis is a complicated, multistep

process . Two of the steps involved with making

a protein are transcription and translation.

Transcription

During transcription, the original DNA code is

rewritten onto a molecule called messenger

RNA (mRNA). mRNA is also made up of a

sequence of nucleotides that differ slightly

from DNA nucleotides . RNA molecules are

very similar to DNA but have a ribose sugar,

are single-stranded and use U, or uracil,

instead of T, or thymine, as one of the four

possible bases .

Translation

During translation, the ribosomes assemble

individual amino acids into proteins . Ribo-

somes will bind to mRNA . Every three mRNA

nucleotides make up a codon—the code for

an amino acid . During translation, transfer

RNA (tRNA) reads mRNA and picks up the

corresponding amino acids . In this way, the

amino acids are linked together in a very

specific combination as dictated by the

sequence of nucleotides on the mRNA .

Short chains of amino acids are called

peptides. Long chains of amino acids are

called polypeptides . Polypeptides fold into

a functional protein .

Hundreds of different types of proteins per-

form specific jobs within and between cells .

Proteins called enzymes put molecules to-

gether and/or break molecules apart . Proteins

called signaling molecules allow cells to relay

messages to one another . Protein receptors

receive signals through a type of communica-

tion known as signal transduction or cell

signaling . Some proteins move substances

into and out of the cell . Structural proteins

give shape to cells and organisms . Proteins

are involved in cellular recognition and identify

different types of cells . Some proteins, such

as antibodies, are involved with defending an

organism against disease .

BIoFaCT

The highest-known number of genes in an organism is around 60,000—for the bacterium that causes trichomoniasis—which is almost three times as many as in the human genome .

The Science8

Biotechnology is based on biology, which is the study of life. The basic unit of life is the cell.

Biologists study the structure and functions of cells—what cells do and how they do it.

Biotechnologists use this information to develop products.

Biomedical researchers use their understanding of genes, proteins and cell parts to pinpoint the

differences between diseased and healthy cells. When researchers know how diseased cells are

altered and when they learn how to affect those alterations, they are better able to develop

innovative medical diagnostics, devices and therapies.

Chapter Three:

How Biology Drives Biotechnology

How Biology Drives Biotechnology 9

Understanding Disease Mechanisms

Early-stage drug research and development

(R&D) may begin with understanding the

underlying biology of a particular disease .

Biotechnology medicines are often created

specifically to address a particular disease

mechanism .

To design and develop new drugs, researchers

must understand the disease mechanisms

involved . Some initial questions researchers

ask to understand the underlying mechanisms

of a disease are: How does a person get

the disease? Which cells become diseased?

Is this disease caused by genetics, and if

so, what genes are turned on or turned off

in the diseased cells? What proteins are

produced—or not produced—in diseased

cells as compared to healthy cells? If the

disease is caused by a pathogen, what is

the interaction between the pathogen and

the person?

Studying disease mechanisms provides

researchers with information that can lead

them to identify targets for the early stages

of the drug discovery process . An under-

standing of fundamental biology may lead

to effective therapies for patients .

Take treatments available for autoimmune

disorders, for example . Autoimmune disorders

occur when a person’s immune system over-

reacts and attacks proteins, cells and tissues

in the body, often leading to inflammation .

Biologists have learned that tumor necrosis

factor (TNF) plays a major role in regulating

inflammation . Researchers know that too

much TNF is produced in autoimmune disor-

ders such as rheumatoid arthritis, psoriasis,

psoriatic arthritis, juvenile idiopathic arthritis

and ankylosing spondylitis . When too much

TNF is produced, excessive inflammation

occurs—and that can be damaging to joints,

skin and other parts of the body . Biotechnology

companies have worked to develop medicines

that inhibit the activity of TNF .

Models for Studying Disease

Researchers often take several different

approaches to creating models for study-

ing a particular disease . One approach is

to obtain samples of diseased cells and

healthy cells and grow them using a method

called cell culture. This calls for cells to be

incubated and fed with specialized growth

media. In culture, the cells do what cells

do—divide and express genes to produce

proteins . By studying how cellular processes

differ between healthy and diseased cells,

researchers hope to come to understand the

mechanism of disease .

Another approach involves studying shared

or similar genes and protein equivalents

in other species . Since all organisms are

made of cells and all cells perform many

similar functions, genes and proteins found in

humans are also found in other organisms .

The functions of human genes have been

revealed by studying parallel genes in

BIoFaCT

The process of analyzing and interpreting biologic scientific data is called computational biology and involves computer science, applied mathematics and statistics .

BIoFaCT

The biotechnology industry is one of the most R&D-intensive industries in the world . The United States is recognized as leading the world in biotechnology R&D .

Autoimmune Diseases Rheumatoid arthritis is a chronic

autoimmune disease that causes

inflammation and tissue damage

in joints and tendons. It can be a

disabling and painful condition

and can lead to substantial loss

of functionality and mobility.

Psoriasis is a noncontagious

disorder that causes red scaly

patches to appear on the skin.

These patches are areas of

inflammation and excessive skin

production.

Psoriatic arthritis is a type of

inflammatory arthritis that affects

up to 30 percent of people suffering

from psoriasis.

Juvenile idiopathic arthritis is

the most common form of chronic

inflammatory arthritis in children.

Ankylosing spondylitis is a

chronic, painful, degenerative

inflammatory arthritis primarily

affecting the spine and causing

eventual fusion of the vertebrae.

Lupus is a chronic inflammatory

disease that most commonly

affects women of childbearing

age. Lupus can affect the kidneys,

joints, blood and skin, among

other organs. Its symptoms can

include rash, fever, aches, anemia

and hair loss.

How Biology Drives Biotechnology10

nonhumans . This approach has added to

our understanding of how specific genes

and proteins direct the functioning of human

cells—both healthy and diseased .

Bioinformatics

Bioinformatics combines biology, computer

science and information technology into one

discipline . The goal of bioinformatics is to

capture, organize and index scientific infor-

mation so researchers can better understand

biology . The challenge for computer pro-

grammers is to design databases that allow

researchers to easily access existing data as

well as submit new data .

The scientific community generates volumes

of biological data daily . Biotechnology

companies use this information to form a

comprehensive picture of normal cell activity

so researchers can better study diseased

cells . This leads to the development of

diagnostic tools, therapies and preventive

medicines . Because of technological ad-

vances in biotechnology, bioinformatics has

evolved to focus on nucleotide sequences,

genes and amino acid sequences .

Biomarkers

Biomarkers are substances that can be

measured and evaluated to indicate (or serve

as markers of) normal biologic processes,

disease processes or biologic responses to

therapeutic treatment and disease intervention .

Historically, biomarkers were physiological

indicators—such as blood pressure or heart

rate . Today, disease may also be detected

using molecular biomarkers such as prostate-

specific antigen, a protein that if elevated,

may indicate prostate cancer .

Once a biomarker is validated (meaning all or

most patients with the disease test positive

for the specific biomarker), it can be used to

diagnose disease risk or presence of disease

or to help doctors determine patient treatment .

Biomarkers also can be used to predict

prognosis—for example, how a disease is

likely to progress if left untreated . Identifying

biomarkers that indicate specific disease is a

crucial step in the R&D process for developing

new biotechnology diagnostic tools .

BIoFaCT

Comparative genomics is the study of genome structure and function among different species . Researchers have obtained complete genomic sequences for the bacterium Escherichia coli (E. coli), the yeast Saccharomyces cerevisiae, the roundworm Caenorhabditis elegans, the fruit fly Drosophila melanogaster, the laboratory mouse and many other organisms for use in comparative genomic studies with the human genome .

KRAS: A Biomarker Breakthrough A biomarker can now help

physicians determine which

patients will not respond to a

certain class of anticancer agents

used to treat metastatic colorectal

cancer (mCRC). Amgen scientists

were the first to show in a phase

3 clinical trial that patients with

mCRC carrying specific cancer-

promoting mutations in a gene

known as KRAS are unlikely to

benefit from a drug designed to

bind to epidermal growth factor receptor (EGFR).

By testing for these mutations,

physicians can now eliminate

anti-EGFR antibodies as a treat-

ment option for patients with

mutated KRAS tumors.

KRAS research is regarded inter-

nationally as an important step

toward personalized medicine

for cancer treatment.

BIoFaCT

Genetic biomarkers are used in developing diagnostic tests for detecting certain genetic diseases . These tests look for DNA fragments identified as causing disease or associated with susceptibility to a disease .

How Biology Drives Biotechnology 11

Researchers study modifications and changes

that occur during cellular processes—such

as protein synthesis—in various disease

states . This can lead to the identification of

biomarkers that signal a change in disease

progression and can be used as a key devel-

opment in drug discovery and testing .

Biomarkers are also used in drug discovery to

determine whether a drug is effective in animal

models and at what doses effectiveness is

reached . Finally, biomarkers can be used in

disease management to determine whether a

drug is having the desired effect and whether

the correct dose is being used . This is very

important because response to a drug can

vary between patients: some require higher

or lower doses for the the drug to be effective .

Proteomics

The entire set of proteins produced by an

organism is called its proteome . Proteomics

is the study of protein structure and function .

Proteins control all aspects of cellular function .

Proteins produced in any specific cell of an

organism can vary with a number of factors

such as time, hormonal change and stress .

Proteomics researchers identify all proteins

involved with protein synthesis and protein

folding . Proteins need to fold correctly into

their 3-D shape in order to correctly function .

Even small structural defects during the folding

process can lead to a number of protein diseases .

Understanding protein structure, function

and interaction within and between cells is

crucial for drug discovery . Proteins are common

drug targets because they are responsible for

most normal cell functions—and malfunctions,

as in the case of disease . A drug target is

the molecule the drug interacts with to bring

about the desired change . This is sometimes

called hitting the target .

Cancer: From Biology to Treatment

Cancer research is a field that has been

at the forefront of utilizing bioinformatics,

biomarkers and protein studies to develop

new therapies that target specific cellular

processes . Cancer biology is a specific field

that explores all aspects of the cancer cell .

Researchers follow cancer pathways and

determine the molecular basis of cancer as

they develop diagnostics and treatments .

Cancer starts with changes in one cell or a

small group of cells that reproduce uncontrol-

lably . In healthy cells, cell division and growth

are tightly regulated . However, cancer cells

keep dividing—without normal checks and

balances—creating greater opportunity for

mutations . A mutation may result from the

environment or could have occurred during

DNA replication . As the cancer cells replicate

and grow, a mass (tumor) is formed . Cells

from the tumor can break away (metasta-

size) and spread through the bloodstream or

lymphatic system to other parts of the body,

creating new tumors .

Traditionally, cancer has been treated with

surgery, radiation and chemotherapy . The bio-

technology industry has contributed significant

advances in cancer treatment by developing

hormone therapies, biologics and targeted

therapies such as monoclonal antibodies .

Types of Cancer Cancer is a general term used

to describe diseases that occur

due to abnormal, uncontrolled

and rapid cell growth. There are

many types of cancer (carcinoma,

lymphoma, leukemia, sarcoma,

melanoma, etc.) that affect many

types of tissue (prostate, skin,

breast, brain, ovaries, lungs, etc.).

Carcinoma is cancer that begins

in the skin or in tissues that line

or cover internal organs. At least

80 percent of all cancers are

carcinomas.

Lymphoma is a cancer of the

lymphatic system—a network

of thin vessels and nodes

throughout the body that helps

to fight infection. Lymphoma

involves lymphocytes, a type

of white blood cell found in the

immune system.

Leukemia is cancer that starts in

blood-forming tissue—such as

bone marrow—and that causes

large numbers of abnormal blood

cells to be produced and to enter

the blood.

Sarcoma is cancer that begins in

bone, cartilage, fat, muscle, blood

vessels or other connective and

supportive tissue.

Melanoma is a cancerous

(malignant) tumor that begins

in the cells that produce skin

coloring (melanocytes). Melanoma

is almost always curable in its

early stages but can be lethal

in later stages.

BIoFaCT

Biotechnology has created more than 200 new biotherapeutics and vaccines, including products to treat cancer, diabetes, HIV/AIDS and autoimmune disorders . The majority of these products are therapeutic proteins .

How Biology Drives Biotechnology12

Biotechnology scientists depend on a wide variety of constantly evolving laboratory techniques and

tools. This section focuses on some of these platform technologies. To understand the industry, it’s

helpful to have some basic knowledge of what goes on in the lab.

Chapter Four:

The Technology

The Technology 13

Restriction Enzymes

Biotechnology employs a process called

genetic engineering, which combines DNA

sequences in order to produce recombinant

proteins as potential therapeutics . The process

utilizes restriction enzymes .

Scientists discovered restriction enzymes

(endonucleases) in bacteria in the 1970s .

They found that these enzymes cut up viral

DNA into small, nonfunctional pieces, thereby

protecting the bacterium from an invading virus .

There are hundreds of restriction enzymes—

each of them recognizing a specific sequence

of DNA called a restriction site . For example,

EcoR1, a restriction enzyme found in E. coli,

recognizes and cuts at the six-base sequence

GAATTC . HaeIII, a restriction enzyme found

in Haemophilus-aegyptius, recognizes and

cuts at the four base sequence GGCC .

All DNA, regardless of where it comes from,

is made up of the same four bases—As, Ts,

Gs and Cs . HaeIII reads any DNA segment

and cuts the DNA every time it encounters

the sequence GGCC . All restriction enzymes

are specific and reproducible, which are two

key characteristics that allow researchers to

utilize restriction enzymes to manipulate DNA .

The counterpart to cutting is called pasting .

DNA ligase is a protein (enzyme) that seals

two DNA segments together in a process

called ligation . The ability to cut and paste

DNA is the basis of genetic engineering .

Recombinant DNA

When segments of DNA are cut and pasted

together, the new DNA is called recombinant

DNA. Recombinant DNA can be inserted into

cells to produce cells with new characteristics .

This genetic altering can include a single-base

(letter) change or multiple gene changes .

Recombinant DNA can be introduced into

a host cell by a vector, which is used to

physically carry DNA into a host cell . A host

cell can be bacterial, yeast, plant, insect

or mammalian .

Common bacterial vectors include plasmids

and phages . A plasmid is a circular unit of

DNA that can be engineered to carry a gene

of interest . A phage is a genetically engineered

virus that injects DNA into bacteria . Cells that

contain recombinant DNA are referred to as

genetically modified, transgenic or transformed

cells . The process is called transformation.

+

BIoFaCT

Daniel Nathans, Werner Arber and Hamilton Smith received the 1978 Nobel Prize in Physiology or Medicine for their discovery of restriction endonucleases . Their discovery led to the development of recombinant DNA technology .

BIoFaCT

Scientists have identified more than 3,800 restriction enzymes, and more than 600 are commercially available for purchase from scientific supply companies .

Transformed CellsTransformation of bacterial cells

for the production of recombinant

proteins usually involves E. coli.

Transformation of animal cells,

called transfection, usually

involves a cell line derived from

Chinese hamster ovary (CHO)

cells. CHO cells were introduced

in the 1960s and remain the most

commonly used mammalian host

cells for industrial production of

recombinant protein therapeutics.

Restriction enzymes are proteins that function as molecular scissors and cut DNA.

The Technology14

The Technology

Recombinant Proteins

Recombinant DNA can be used to produce

recombinant proteins. The host cells use

the new DNA information and their own

cellular machinery to produce the protein

encoded by the recombinant DNA .

When recombinant proteins are produced for

use as human therapeutics, host cells must

be grown in large quantities so that enough

recombinant protein is produced to meet

demand . The recombinant protein is isolated,

purified and analyzed for activity and quality

before it goes to market .

Producing a protein with the proper order

of amino acids isn’t always the whole story .

Sometimes further processing is required to

yield an active or fully functioning protein .

Many human proteins are glycosylated,

meaning, they have a particular pattern of

sugar molecules linked to them . If a protein

is translated but not correctly glycosylated,

it may not function properly .

Adding a phosphate group—a process

known as phosphorylation—can act as an

on-switch, allowing the proteins to become

active proteins . Other biochemical functional

groups may be added to the protein to allow

it a larger range of function .

Recombinant proteins for therapeutic use

include vaccines, hormones, monoclonal

antibodies and hematopoietic growth factors

for the treatment of cancer, AIDS, aller-

gies, asthma and many other conditions .

The number of recombinant proteins has

increased greatly in recent years as the

technology used for their production and

purification has advanced .

Cell Culture

Cell culture is the technique of growing cells

in the laboratory under controlled conditions .

Growing large quantities of transformed cells

is a major step in the process of producing

recombinant protein products . Both

transformed bacterial cells and transformed

animal cells are used in this process .

Simple proteins can be produced using

recombinant DNA technology in bacterial cell

cultures . Typically, more-complex proteins

that are, for example, glycosylated are made

in animal cell cultures .

During cell culture, cells are grown in petri

dishes or flasks containing liquid media . Culture

media provides all the nutrients necessary

for cell growth . The cultures are grown in an

incubator that maintains the appropriate

temperature and environment, often requiring

certain gas mixtures of oxygen and carbon

dioxide . It is very important to maintain

specific conditions for cultures, because

variation in these conditions may affect

the proteins expressed and, therefore, the

product that is produced .

In the process of commercial production of

proteins, cell cultures must be scaled up to

produce enough protein to meet demand .

Since only a limited number of cells can be

grown in small petri dishes or flasks, the cell

culture can be transferred to large containers

called bioreactors, which are involved in the

manufacturing process .

BIoFaCT

The FDA approved human insulin in 1982—the first medicine made via recombinant DNA technology .

15

Research Tools

Biotechnology professionals depend on

leading-edge laboratory equipment . Here are

a few pieces used in genetic engineering:

Thermocycler

Polymerase chain reaction (PCR) is a

technique that replicates DNA in a machine

called a thermocycler. PCR is a series of

cycles that takes a small amount of original

DNA and exponentially copies, or amplifies, it .

Each three-step cycle doubles the amount of

DNA present . It’s like a photocopier for DNA .

A single piece of DNA can be turned into

millions of copies of the same DNA piece .

PCR enables researchers to make enough

DNA to work with in the lab . There are many

applications for PCR, including producing

enough of a DNA sequence or gene for use

in creating recombinant proteins .

Gel Electrophoresis

Gel electrophoresis is a technique commonly

used in the laboratory for analyzing DNA

fragments . Gel electrophoresis allows DNA

fragments to be separated within a gel . A gel

electrophoresis apparatus holds the gel and

allows electricity to run through it . Each DNA

fragment is negatively charged, causing it to

migrate toward the positive pole of the gel

apparatus . Larger fragments of DNA move

more slowly than smaller fragments because

they encounter resistance from the gel matrix .

There are different types of electrophoresis .

Each is based on the type of separating

material used, gels being one type, and the

types of molecules being separated . DNA,

RNA and proteins can all be separated using

electrophoresis in an appropriate apparatus .

Gel electrophoresis has many applications in

both clinical and research labs . One common

use is for verifying PCR products—that is,

checking to see whether the reaction generated

the correct DNA fragment .

DNA Microarrays

A DNA microarray (also called a gene chip)

is a small piece of glass or silicon divided

into thousands of sections in a grid pattern .

Each section has a single-stranded gene

fragment corresponding to either a healthy

or diseased gene .

DNA from an individual is separated into

single strands and tagged with a fluorescent

dye . The tagged DNA is washed over the

microarray . The individual’s DNA binds to

any complementary DNA sequences on the

slide, if they are present, to become double-

stranded DNA .

With the aid of a computer, the double-stranded,

fluorescently tagged DNA spots can be located

and measured . The individual’s DNA must

match the attached gene fragment exactly

to bind, thereby indicating the presence of

healthy or diseased DNA .

BIoFaCT

Kary Banks Mullis developed the polymerase chain reaction in 1983 and received the 1993 Nobel Prize in chemistry for the invention .

Gel electrophoresis.

The Technology

- +GelWell

16

Microarrays are powerful tools allowing

researchers to analyze thousands of genes

in one test . Microarrays are used for genetic

testing, for comparison of genetic information

from different individuals or species, and in

the discovery of potential drug targets .

Microarrays can also be used for researching

and identifying genes of interest to be used

with recombinant DNA technologies . Other

microarrays include protein, tissue, chemical

compound and antibody microarrays—all of

which allow researchers to analyze thousands

of data points at one time .

The Technology

Microarray.

BIoFaCT

Microarrays are capable of generating so much data that special microarray data-bases are needed for storage, searches, analysis and interpretation . There are both public and private microarray databases .

17

During drug discovery, scientists search for molecules—either chemical or biological agents—that

could alter a disease pathway. As part of the discovery process, they specifically look for ways to

change one or more molecular or cellular processes that occur in the affected cells of a diseased

tissue or organ.

Chapter Five:

Drug Discovery

Drug Discovery18

Drug Discovery

Chemical LibrariesIn the 1990s, chemists developed

huge libraries of chemical

compounds—thousands, even

millions, of chemicals with dif-

ferent structures used to screen

for new drugs. These libraries are

often proprietary and constructed

by a company explicitly to support

its drug discovery programs.

Initial screening of drug candi-

dates is relatively simple. Once

potential drug candidates or leads

are identified, more-complex

assays are used at subsequent

levels of screening. The sources

of the compounds found in the

drug candidates are often natural

products (from microbes, plants

and simple marine life) or chemi-

cal compounds synthesized by an

organic chemist.

Combinatorial chemistry increases

the potential of chemical libraries

by synthesizing larger, more

complex chemicals or chemically

related molecules from common

chemical structures. Combinatorial

chemistry for small-molecule

drugs includes the synthesis

of large organic molecules by

adding together smaller organic

molecules, often with improved

product results or lessened

product side effects.

Drug Discovery

Initiating Drug Discovery Research

An early step in the drug discovery process

is to identify an unmet medical need . What

is known about the disease? What are the

current treatment options, if any? Does the

company have the expertise, technology

and financial resources to solve the prob-

lem? Potential competitors and barriers, such

as regulatory constraints, are also taken into

consideration .

Target Discovery

After identifying an unmet medical need and

deciding whether it fits within the company’s

portfolio, scientists look very closely at the

biology behind the disease . Where can they

intervene, and what options do they have for

intervention? Since the human body is an

extremely complex system, scientists have

to carefully choose the target .

A target is a molecule that plays a critical role

in a disease . Scientists estimate that about

8,000 known therapeutic targets exist today .

Targets can be secreted factors, cell surface

receptors or signaling pathways within a cell .

The goal is to develop a drug that affects

a target in a way that interferes with the

disease process . It’s also very important to

ensure that the potential benefits of a drug

are appropriately weighed against any risks

such as possible side effects .

Different targets respond to different therapeutic

approaches . To select a target, scientists will

ask, “What are the differences between healthy

and diseased cells?”

Ultimately, disease processes take place at

the molecular level . There are various causes

of diseases . In inherited diseases, a difference

in the expression or in the sequence of genes

results in abnormal functioning of a person’s

cells . Sometimes this leads to a target being

present in excess; other times it could be

deficient or missing . So the scientists will

need to decide if the goal will be to block the

target or to enhance or replace it in order to

restore healthy function . For a disease caused

by an external pathogen, such as a virus or

bacterium, the pathogen produces molecules

that can damage the host organism’s cells .

Moreover, the pathogen will, itself, display

molecules in the infected individual that are

not present in a healthy person . The goal in

target discovery is to identify those differ-

ent molecules . This can be done using a

variety of technologies such as microarray

experiments, protein electrophoresis, mass

spectrometry (MS), DNA sequencing and

computerized imaging .

While this sounds straightforward, target

discovery is often difficult and may take years

to complete . Why? Cells and cell-to-cell

interactions are very complex . There may be

one or more mechanisms of the disease and

many points in the mechanism at which to

intercede . Moreover, the difference between

healthy and diseased cells can be too minute

to easily detect, or a method able to detect

the difference may not yet have been invented .

BIoFaCT

According to the National Center for Health Statistics, the top five diseases causing death in the United States in 2005 were heart disease, cancer, stroke, chronic lower respiratory disease and diabetes .

BIoFaCT

Researching the genetic and molecular basis of a disease is called studying the mechanism of disease .

19

The complexity of the body’s response also

means scientists could see a difference in

the expression of hundreds of genes without

being able to determine which ones were

critical to the disease .

Target Validation

Once scientists identify potential targets, the

next step is to validate them . Target validation

has two components . The first is to show

that the target molecule actually plays a role

in the disease . The second is to confirm the

target is a candidate for therapeutic interven-

tion: Can a safe and effective drug be made

against the target? Scientists complete this

second component of target validation before

the drug enters human testing .

There are a number of ways to validate a

target, and the process must take into con-

sideration time, cost and technology . At the

simplest level, the concept of target valida-

tion is to use the target to create the disease

in a sample of healthy tissues and then block

the target to restore the healthy condition .

This is done in cell culture or animal models .

The trick is to select a model that is repre-

sentative and will work . Sometimes people

who are born without certain functional

molecules express a specific disease type .

Studying biological samples taken from such

human subjects provides another means of

validating a target .

Examples of target molecules include recep-

tors, enzymes, ion channels, growth factors,

cytokines and DNA binding proteins . The

common thread among these targets is that

they are often involved in signal transduction

processes in and among cells . Signal trans-

duction pathways control cellular processes

such as division, differentiation, protein synthe-

sis and programmed cell death (apoptosis).

Initial studies are often done in cell culture . If

cell culture studies are positive, a next step

is to use an animal model .

Sometimes a suitable animal model has to be

created to validate a target . Sometimes the

target doesn’t exist in an animal model or may

not mimic the human disease state . Some-

times the drug candidate is so specific to

humans, it won’t recognize the animal model’s

target or the animal will mount an immune

response that blocks any therapeutic effect .

For example, Alzheimer’s disease occurs only

in humans, and only recently have mouse

models been developed to mimic the disease .

Scientists also look at what other effects the

drug candidate may have within preclinical

(both cell culture and animal) models . Some-

times the target is expressed on other cells or

tissues besides those directly involved in the

disease . What happens to those cells and

tissues in the presence of a drug candidate?

Does a drug candidate adversely affect other

cells or tissues? Does it raise an immune

response; stimulate other, similar targets; or

otherwise present any concerns about toxicity?

Preclinical work helps support later human

trials that may occur if the drug candidate

continues to show promise . Even if the drug

gets marketing approval after successfully

completing the necessary phases of human

Drug Discovery

Cell Receptors and Ion ChannelsThe most common drug targets

are cell receptors—proteins on

or inside a cell to which a specific

signaling molecule can attach.

These signaling molecules can be

hormones, neurotransmitters, pharmaceutical drugs, toxins or

even infectious agents. When

signaling molecules attach to

the receptor, a physical change

occurs that initiates a specific

cellular response.

Other common drug targets are

ion channels, proteins that form

pores in the membranes that

surround cells, and enzymes—

proteins that increase the rate of

specific chemical reactions.

BIoFaCT

Recently, scientists have begun using computer simulation to model drug-target interactions to guide drug discovery .

20

Drug Discovery

Choosing the Right Tool for the TargetDesigning a targeting strategy

usually comes down to a choice

between a small-molecule drug

and a biologic (most often a

recombinant protein or antibody).

Each has its particular advantages

and disadvantages.

Small molecules can usually cross

cell membranes and enter cells,

allowing them to be used for

targets inside cells. Biologics usu-

ally cannot cross cell membranes,

restricting their use to targets on

the surface of, or outside, cells.

Small molecules have good

specificity for their targets, but

recombinant antibodies generally

have extremely high specificity,

meaning, fewer adverse reactions

for the patient. Small molecules

have variable half-lives, which is a

measurement of how long a drug

stays active in the bloodstream

or in its target tissues. Biologics

often have much longer half-lives,

partly because they are modeled

on real biological molecules. This

means patients don’t have to

take as many doses of a biologic,

which may result in better patient

adherence to therapy.

Biologics usually need to

be injected, whereas small

molecules can be taken orally.

Small molecules can often cross

the blood-brain barrier, but

biologics usually cannot, which to

date has limited their usefulness

for treating diseases of the brain

such as psychiatric disorders and

neurodegenerative diseases.

trials, safety surveillance will continue once the

drug has reached the larger patient population .

Scientists will continue to answer safety

questions throughout the life of a drug .

Screening

High-throughput screening is a process

that combines robotics and data processing

to rapidly identify the compounds, antibodies

or genes that modulate a particular biomo-

lecular pathway . Large batches of potential

drugs are tested for binding activity or bio-

logical activity against target molecules .

Once a candidate disease is identified, a

company’s research lab develops a testing

method (assay) to determine or measure

the pharmacological activity of hundreds to

hundreds of thousands of molecules .

The assay measures the estimated potential

of a molecule to block or stimulate a target .

What’s being measured could be as simple

as the ability of the drug candidate to kill

cancer cells in culture or as complex as mea-

suring its ability to inhibit an enzyme involved

in a disease . Generally, the more complex the

assay, the more relevant the information—

but the higher the cost of the assay and the

longer it usually takes to get data .

Of the molecules that score a hit—that is,

a positive result that appears to have a

therapeutic potential—some are identified

as lead molecules due to their more druglike

properties (solubility, permeability, stability,

etc .) . Once a drug candidate is identified,

scientists may attempt to optimize its ability

to fight disease by changing its molecular

structure through combinatorial chemistry

for small molecules or protein engineering

for large molecules .

Drug Design

The design approach to drug discovery starts

with scientists understanding the genetic and

molecular base of a disease and using that

information to select a specific therapeutic

target . Drugs are then designed to interact

with the target . Through rational drug design,

scientists seek to develop a drug that is highly

specific to a particular target in a disease in

hopes of achieving a better therapeutic out-

come with potentially fewer side effects .

Scientists can learn more about the structure

of the target by using imaging technology

such as X-ray crystallography . 3-D structural

information about a target enhances drug

design strategies .

Considerations in designing a therapeutic

agent depend on both the nature of the target

and the capabilities of the company . If the tar-

get is on the exterior surface of the cell mem-

brane or is secreted, protein therapeutics such

as monoclonal antibodies or peptides can be

used . If the target is on the interior of the cell,

only drugs that can cross the cell membrane,

such as small molecules, can be used .

When designing a drug candidate, scientists

must keep in mind the intended method of

drug delivery and determine whether the drug

will be a pill swallowed, a liquid injected, a

spray inhaled or something else .

Drug Discovery 21

After the lengthy process of drug discovery (identifying a target and validating a drug candidate),

the process of drug development is still far from complete. Drug development includes the safety,

efficacy, formulation and manufacture of the drug. Typically, safety testing begins with a series

of experiments called preclinical studies. If these studies predict the drug candidate to be safe,

testing begins in humans in a series of studies called clinical trials.

Chapter Six:

Drug Development

Drug Development22

Drug Development*For more information on Amgen’s commitment to ethical use of animals in research, visit www .amgen .com/science/ethical_research .html .

Preclinical Studies

Preclinical studies are tests that take place

in a scientifically controlled setting using cell

cultures and animals as models . The goal of

preclinical studies is to predict what the body

does to the drug candidate (pharmacokinetics),

what the drug candidate does to the body

(pharmacodynamics), and whether the

drug candidate may pose potential health

hazards or toxic side effects .

Pharmacokinetic testing provides data to

answer questions such as: How is the drug

absorbed and transported? Which cells

and organs are affected? What enzymes

in the body break down the drug, and how

fast does this occur? How is the drug or its

metabolites (breakdown products) eliminated

from the body? Pharmacodynamic studies

examine dose-response effects and often

monitor biochemical and physiological changes

(such as enzyme activities, heart rate, blood

pressure and body temperature) in the test

subject . Pharmacodynamic testing, which

shows what the body does in response to

the drug, is used to answer the question:

Is the drug harmful or toxic to cells or organ

systems? Toxicology studies address the

potential of the drug or its metabolites to kill

or damage cells and organs, cause cancer or

cause reproductive problems, including birth

defects or sterility . Pharmacokinetic and

pharmacodynamic studies are used together

to reach the goal of preclinical studies, which

then answer the question: Is the drug safe?

In the United States, preclinical studies must

be conducted under FDA guidelines known

as current Good Laboratory Practice . Many

other countries follow global harmonized

regulatory guidelines as well .

Information from these studies is vital . It allows

researchers to estimate a safe dosage level

for humans in phase 1 clinical trials . Although

drug companies are required to submit animal

model data to regulatory agencies as part

of the drug approval process, companies are

taking steps to reduce the number of animals

used in testing because of ethical concerns

and the cost associated with facilities .

In the United States, institutes that conduct

research involving animals and that receive

federal funding must have an Institutional

Animal Care and Use Committee (IACUC) .

This committee reviews research protocols

and evaluates the care laboratory animals

receive . The IACUC is responsible for making

sure labs comply with the Animal Welfare Act .

Animal models greatly enhance scientists’

ability to test the effectiveness and safety of

new drug candidates . In target validation,

researchers may use knock-out mice and/or

knock-in mice to validate a target . Knock-out

mice are genetically altered to remove mouse

versions of human disease genes . Human

disease genes can also be knocked in to

create mouse models with human diseases like

cancer, diabetes, Alzheimer’s and Parkinson’s .

Drug candidates are tested on these mice,

enabling researchers to check for adverse

side effects before giving the candidate drug

to humans .

Initially, many studies of drug safety and

toxicity are done using cell lines. Cell lines

are engineered to express genes that are

often responsible for adverse reactions . The

creation of cell line models has decreased

the number of animals needed for testing

(reducing cost and time) and helps accelerate

the drug development process .*

23

Drug Development

How Is Dosage Determined? There are two types of phase 1

dosage studies: SAD studies and

MAD studies.

SAD: Single-Ascending-Dose Studies A few volunteers are given a

small dose of the investigational

new drug and observed. If there

are no adverse reactions, another

group is given a slightly higher

dose. This is repeated as many

times as needed until volunteers

start to exhibit intolerable side ef-

fects. At these dosage levels, the

investigational new drug is said to

have reached maximum tolerated

dose (MTD).

MAD: Multiple-Ascending-Dose Studies The same volunteers receive

higher and higher doses of the

investigational new drug until

the dosage reaches a certain

level. Samples of body fluids are

collected with each increase in

dosage level to understand

how the body processes the

investigational new drug.

Phase 1 clinical trials are usually

conducted in an inpatient clinic

where full-time staff can observe

the study subjects.

If preclinical trials provide sufficient evidence

that a drug candidate is safe, companies

submit an Investigational New Drug (IND)

application to the FDA . After the FDA approves

the IND, companies can begin phased clinical

trials in humans .

Clinical Trials

Clinical trials are tests designed to determine

the safety, proper dosage, efficacy, adverse

reactions and long-term-use effects of a new

drug in human subjects . Clinical trials taking

place in humans are conducted under global

harmonized guidelines, such as the FDA’s

current Good Clinical Practice (cGCP),

which protects the rights and ensures the

safety of human test subjects and follows the

U .S . Code of Human Research Ethics .

Clinical trials are conducted in three successive

phases—1, 2 and 3—and test progressively

larger numbers of humans in each phase . Each

phase has a different purpose and helps re-

searchers answer different questions . If a phase

is successful, the drug candidate proceeds to

the next phase . If unsuccessful, clinical trials

are halted, the drug is suspended and the

sponsor company returns to the discovery

phase to look for another drug candidate .

Clinical trials are conducted at different testing

sites . It takes several years to complete all

three clinical trial phases .

Clinical trials are often managed by a contract

research organization (CRO), which is an

independent organization . The CRO is

responsible for all the data management and

communication between the sponsor com-

pany and physicians overseeing the clinical

trials . The CRO also ensures that the study

volunteers understand and accept the risks

involved in the clinical trials and that cGCP

guidelines are followed .

Phase 1

Phase 1 trials represent the first time an

investigational new drug is tested on humans .

The goal is to evaluate the drug’s safety, tol-

erability and safe dosage range . The testing

group is often small, ranging from 20 to 50

volunteers . These are usually healthy volun-

teers who do not have a disease . However,

sometimes patient volunteers will be ac-

cepted into a phase 1 clinical trial, particularly

when testing oncology therapeutics . Usually

these patients have been unsuccessful with

available treatments or have few treatment

options, or the drug’s potential side effects

are too risky to involve healthy subjects (such

as using some chemotherapeutic agents) .

Phase 2

The goal of phase 2 trials is to determine

the efficacy and safety of the investigational

new drug among a larger group of patient

volunteers—usually 100 to 300 people .

A patient volunteer is someone who has the

disease the drug is intended to treat . Some

companies divide phase 2 trials into phase

2A (to assess dosage) and phase 2B

(to assess efficacy) . Most investigational

new drugs fail during this stage because

of efficacy and/or safety issues .

BIoFaCT

A crucial component of initiating a clinical trial is recruiting study subjects who agree to participate and sign a document called informed consent . Potential subjects must be informed about all aspects of the study before they decide to participate . Participants can withdraw their informed consent at any time .

24

Drug Development

Phase 3

The goal of phase 3 trials is to confirm the

effectiveness of the investigational new

drug and compare it with placebos or

therapies already available on the market .

To do this, hundreds or thousands of patient

volunteers are tested . Phase 3 trials are the

most expensive and time-consuming, lasting

for a couple of years or longer to establish

long-term safety .

Once phase 3 is successfully complete, the

sponsor company files a new drug or biolog-

ics application with the country’s regulatory

agency . In the United States, the company

would file a New Drug Application (NDA) for a

small-molecule drug or a Biologic License

Application (BLA) for a large-molecule drug

with the FDA . If the governing regulatory au-

thority (the FDA in the United States or the

European Medicines Agency, known as the

EMEA, in Europe) approves the drug, the

sponsor company is permitted to market and

sell the product in the country or countries

regulated by that authority . The final manufac-

turing of the drug—or large-scale production—

must take place in a facility that meets the

country’s strict guidelines, such as the FDA’s

current Good Manufacturing Practice (cGMP),

to ensure safety and purity of the product .

Phase 4

Phase 4 trials occur after an approved drug is

on the market . A goal is to monitor the drug’s

safety and efficacy when utilized in a normal

medical setting in a population of patients

that could number in the millions . Sometimes

adverse reactions, which were not seen in

a comparatively small cohort of patients

(3,000 patient volunteers as compared to

millions), are discovered in larger and more

diverse populations . If an adverse reaction is

discovered, the drug may be withdrawn from

the market . Either the sponsor company can

voluntarily withdraw the drug or a regulatory

body can pull the drug from the marketplace .

After further testing, the drug may or may not

be reinstated .

The stages in product development, or

product pipeline, take, on average, 10 to

15 years to complete . Most investigational

drugs do not make it . Out of every 1,000

potential new drugs in discovery, only one

will make it to approval .

Study DesignsLate-stage trials often include

a double-blind randomized

controlled test. In this type of

study, neither the patient volun-

teer nor the researcher knows

which volunteer belongs to the

control group or the experimental

group. Each patient volunteer is

randomly placed into one of the

groups. A third party keeps this

documentation and releases it

only after the study is over.

BIoFaCT

One of the largest challenges associated with clinical trials is the shortage of study subjects .

25

The manufacturing of biologics is complex, since most are proteins—large molecules often variable

in structure and sensitive to environmental conditions. The manufacturing of biologics has become

a science that can be summarized in four key steps: producing the master cell line, growing cells

and producing protein, isolating and purifying protein from cells, and preparing the biologic for

patients. The whole process from creating the master cell bank to preparing the biologic for

patients can take years and cost hundreds of millions of dollars.

Chapter Seven:

Scale-Up and Manufacturing

Scale-Up and Manufacturing26

Cell BanksCell banks involve a two-tiered

frozen cell banking system: a

master cell bank (MCB) and a

working cell bank (WCB). Scien-

tists use one vial of cells from

the MCB to create the WCB. Once

established, the WCB is used to

produce batches of product in

the scale-up process. Working

from the same stock of cell line

reduces the chance of mutations

associated with serial cultures.

The MCB is a reserve of cells

that scientists use only when

absolutely necessary. To protect

the integrity of the cell lines,

companies store their cell banks

in two or more locations within

their facilities and in one location

off-site.

Growth vessels vary in size:• Flaskshold5mL

• Spinnerflasksorrollerbottles

hold 50 to 200 mL

• Benchtopbioreactorshold

5 to 20 liters

• Pilotscalebioreactorshold

50 to 200 liters

• Productionvesselshold

20,000 liters or more

Using R&D Specifications

During the R&D phase, researchers develop

the initial production methods on a small scale .

They also determine the drug’s final formula-

tion, or physical form, for clinical trials—for a

biotechnology medicine, usually an injection

or infusion . Using all of the R&D data from

these production steps, companies devise

large-scale production methods to produce

enough of the product for the intended market .

The scale-up and manufacturing process must

adhere to cGMP guidelines to ensure product

safety and purity .

Common Cell Lines

Many biotechnology products are proteins

that must be produced by cells grown in

culture . Chinese hamster ovary (CHO)

cells, nonsecreting (NS0) cells (pronounced

“NS zero”) and E. coli are cell lines used for

production of biotherapeutics, especially

monoclonal antibodies .

There are a number of reasons to use these cells .

Both CHO and NS0 cells synthesize proteins

much like human cells do . Both are immortal

cell lines, meaning, they should be able to

grow and produce product forever . Researchers

are well versed in their optimal culture conditions .

Both cell lines have generally regarded as

safe (GRAS) status for therapeutic protein

production . NS0 cells have the additional

advantage of being programmed to produce

antibodies, but they do not make or secrete

any of their own antibody protein .

Other cell lines can be used and may be

more suitable . The selection of the cell line

depends on the expertise of the company,

the properties of the cell and the regulatory

requirements .

Scale-Up Process

The scale-up of a cell culture process can be

very difficult and time-consuming, taking as

long as several months before researchers

can obtain a product . The entire process of

producing a biotech product from start to

finish is often called a campaign and is usually

divided into two main parts: upstream and

downstream. Upstream processes involve

production of the protein product, most often

by using cells (microbial, insect or mammalian)

growing in culture . Downstream processes

include the recovery, purification, formulation

and packaging of the protein product .

Upstream Phase

Upstream processing begins with the cells

that scientists create or engineer to make

the protein product . Once the desired cell

line is made, it is cryopreserved: scientists

freeze a large number of vials of the cells to

create a cell bank. To begin a campaign,

scientists remove and thaw a vial of cells

from the cell bank and initiate a cell culture

in a flask containing a small volume of

growth media . The initial volume of media

can be as little as 5 mL . The media provides

the nutrients and the optimum environment

for cells to survive .

Scale-up is done by gradually transferring the

growing cells into successively larger growth

vessels containing larger media volumes .

The cells are constantly dividing as long as

the growth environment remains favorable .

Therefore, more and more cells are present

BIoFaCT

The American Type Culture Collection (ATCC) is a private, nonprofit resource dedicated to the collection, preservation and distribution of authentic cell lines and other biologic materials . (www .atcc .org)

Scale-Up and Manufacturing 27

with each step . The greater the number of

cells, the more protein product is generated .

Scale-Up Monitoring

The goal of the scale-up process is to grow

cells as quickly as possible and to produce

as much protein product as possible .

Using the same assays or testing methods

used in the initial R&D stages, scientists

measure cell viability and concentration,

product concentration and product activity

at each incremental scale-up stage for

monitoring purposes .

Lab technicians monitor and control the

physical environment in which cell cultures

grow . They do this manually in the initial

scale-up steps to optimize growth param-

eters such as temperature, pH, nutrient

concentration and oxygen level .

The monitoring process is automated once

the cell culture is large enough to be grown

in bioreactors .

It is crucial during the scale-up, fermentation

and manufacturing stages that technicians

monitor and test the cultures for contamination

by bacteria, yeast or other microorganisms .

Any contamination of a culture ruins the entire

batch of product and costs a company money

and time . Technicians follow very strict protocols

for maintaining aseptic conditions at all times

during the scale-up and manufacturing stages .

Quality Control and Quality Assurance

Quality control (QC) and quality assurance

(QA) departments are responsible for all of

the monitoring that is crucial to the success

of the scale-up and manufacturing stages of

product development . The QC department

assures product quality and testing during

the product development stages well before

the product is at the stage of marketing,

ensuring that the scale-up and manufacturing

processes meet certain standards . The QA

department is usually responsible for meeting

and reporting quality objectives .

Downstream Phase

In the downstream phase of manufacturing,

the protein product is isolated from the cells

that produced it . Proteins found inside the

cell (intracellular proteins) require special

protocols to extract them for purification .

Usually this involves bursting the cells open

to release the protein product, which then

has to be purified away from the other com-

ponents that were inside the cell . Proteins

found outside of the cell (extracellular

proteins) can be easier to isolate .

After harvesting the protein product, the

next step is clarification. This is where

scientists separate the protein from cellular

debris . Then they apply the protein solution

to a series of chromatography columns to

obtain a pure protein product . Purification of

protein mixtures by column chromatography

separates proteins based on physical and

chemical properties such as size, shape or

charge (+ or –) . Additional purification steps

remove any residual DNA and deactivate any

viral particles that may be present .

Researchers verify the isolation and

purification of the protein product through

confirmed testing protocols . The protein

product is then formulated according to

the R&D specifications and packaged for

use by physicians and patients .

Scale-Up and Manufacturing28

Biotechnology Medicines

The biotechnology industry uses advanced technologies to apply cellular and molecular biology

to create new, beneficial products. Medical biotechnology products are used to treat or prevent

diseases. These products include therapeutic proteins, monoclonal antibodies, vaccines, allergy

immunotherapy products, blood components and tissues and cells for transplantation.

Chapter Eight:

Biotechnology Medicines

29

Biotechnology Medicines

Therapeutic Proteins

Scientists use recombinant DNA technology

to make therapeutic proteins, often referred

to as biologics . Biologics are used in such

fields as oncology, rheumatology, immunology,

endocrinology and virology . Approximately

50 recombinant therapeutic proteins are

approved for clinical use and are currently

marketed, and hundreds more are undergoing

clinical trials . Some biologics have been in

use for more than 20 years and are considered

standard therapy .

Doctors have long used therapeutic proteins to

replace or supplement patients’ natural body

proteins—especially when natural protein

levels are decreased or lost due to disease .

Some recombinant proteins are versions of natural

body proteins, and other versions are not exact

versions but produce similar effects in the body .

Vaccines

Vaccines stimulate the immune system and

provide protection against particular diseases .

The first vaccines were made with inactivated

(killed) or weakened virus unable to reproduce

in the body but sufficient to provide immunity

upon future exposure to the live virus .

Vaccines are also created with recombinant

proteins . Scientists use genetic engineering

to create recombinant vaccines by inserting

genes for desired antigens into a vector . A

vaccine vector, or carrier, is a weakened virus

or bacterium into which harmless genetic ma-

terial from another disease-causing organism

can be inserted . Typically, the body recognizes

antigens as foreign, and white blood cells will

attack them . Recombinant vaccines, however,

do not cause disease but do have the antigen,

thus tricking the body into thinking it is being

attacked by a pathogenic virus . Recombinant

vaccines are safe and easily grown and stored .

Antibodies

A major area of biologics is the production

of humanized or fully human antibodies .

Antibodies can attach to antigens found on a

pathogen and flag the pathogen for destruction

by the immune system . Antibodies also can

attach to proteins on immune cells that are

involved in autoimmune responses in diseases

like rheumatoid arthritis and multiple sclerosis .

Humanized antibodies are engineered to be

mostly human to avoid problems with rejection .

Fully human antibodies are derived from human

cells or human antibody genes .

Peptibodies

Peptibodies are engineered therapeutic fusion

proteins with attributes of both peptides and

antibodies but are distinct from each, and bind

to human targets .

Diagnostics

In addition to recombinant proteins being used

as biologic drugs, scientists use recombinant

DNA technology to produce a number of

diagnostic tests for diseases, including tests

for hepatitis and AIDS . In fact, scientists

commonly use recombinant protein antigens

as diagnostic reagents in enzyme-linked

immunosorbent assays (ELISAs) for the

detection of infectious agents such as Severe

Acute Respiratory Syndrome (SARS) .

Monoclonal AntibodiesThough monoclonal antibody

technology was invented in the

mid-1970s, it took 20 years

before the technology showed

its true potential. The first

experimental monoclonal antibod-

ies developed in mouse models

were ineffective because the

human immune system rejected

mouse antibodies as foreign. The

subsequent development of first

humanized, then fully human,

antibodies has enabled the suc-

cessful use of this breakthrough

technology in fighting cancer and

other serious illnesses.

Biotechnology Medicines30

Future of Biotechnology in Healthcare

Biotechnology can offer patients more and better healthcare choices. New, innovative diagnostics

and therapies are changing how some human diseases are prevented and others are treated. This

monumental healthcare shift is in its early stages, with novel medicines, diagnostics and technologies

in development that hold great potential to improve patients’ lives.

Chapter Nine:

Future of Biotechnology in Healthcare

31

Future of Biotechnology in Healthcare

Personalized Medicine

Personalized medicine is the concept that

patients should be treated with therapies and

medicines based specifically on each patient’s

unique genetic makeup, for optimal results .

Currently, the practice of medicine is based

on standards of care that are determined by

averaging responses across large groups of

people . Personalized medicine is a new para-

digm that proposes to manage a patient’s

disease based on the individual patient’s spe-

cific characteristics, including age, gender,

height, weight, diet, genetics and environ-

ment . Genetic testing is beginning to allow

the development of genomic personalized

medicine—medical care based on a patient’s

genotype or gene expression profile .

Pharmacogenomics

A major movement in healthcare is phar-

macogenomics. Pharmacogenomics takes

advantage of the fact that individuals have

unique genomes representing their genetic

makeup . Each genome is likely to react differ-

ently to a particular drug and dose amount .

The challenge is to identify which drug and

which dose will work most optimally for

each person or for groups of individuals who

share similar genetics . By understanding a

patient’s genetic makeup, a physician can

better prescribe a drug and dose level that will

optimally work to combat a particular disease .

Advances in DNA technology are the keys to

both pharmacogenomics and personalized

medicine . These advances allow for testing

and identifying an individual’s unique genetic

makeup and then comparing those differenc-

es with the population at large . Knowledge of

the human genome, variations of the genome

among individuals and variations of the encod-

ed proteins produced enables researchers to

develop medicines that address the individual

needs of each patient . Pharmacogenomics

and personalized medicine promise to improve

clinical trials for new drugs, advance screening

technology for diseases and result in more-

effective individualized healthcare and advances

in preventive medicine .

Genetic Testing

The biotechnology industry has brought about

vast improvements in testing and diagnosis for

genetic diseases . The discovery of single-

nucleotide polymorphisms (SNPs)—single-

nucleotide changes in the DNA sequence—

was one of the major breakthroughs in genetic

testing . SNPs (pronounced “snips”) represent

one of the most common forms of genetic

variation among individuals . When a SNP

occurs in a gene sequence that encodes for

a specific protein, it may change that protein

and cause a disease or increase a patient’s

susceptibility to a disease . Utilizing technology

to detect SNPs allows for more-accurate

diagnosis of genetic diseases and therefore

facilitates treatment decisions . Genetic testing

provides patients with both an understanding

of possible risks for certain diseases and

possible opportunities for prevention .

BIoFaCT

Molecular diagnostic tests analyze DNA, RNA or protein molecules to identify a disease, determine its course, evaluate responses to therapy or predict individual predisposition to a disease .

BIoFaCT

Approximately 10 million SNPs have been identified in the human genome .

32

Gene Therapy

Gene therapy is an emerging area of applied

genetics that utilizes recombinant DNA tech-

niques . In this case, the recombinant DNA

molecules themselves are used for therapy .

Gene therapy involves inserting genes,

created by recombinant DNA technology,

into the cells and tissues of patients to treat

their diseases . Scientists are studying gene

therapies for a number of inherited human

diseases involving defective genes . The idea

is to replace them with new, functional genes .

Since the first clinical trial was initiated in

1990, gene therapy research has expanded

greatly, with an increasing number of human

trials . The field, still in experimental stages,

focuses its efforts on patients with severe

and life-threatening diseases who usually

have few treatment options or who have

failed all available therapies .

Stem Cells

Stem cells are unspecialized cells that can

renew themselves indefinitely to produce more

stem cells . They can mature and develop

specialized functions or differentiate under

specific growth conditions . Stem cells eventually

differentiate to form all of the different types of

cells that make up the body . The broad potential

of an undifferentiated stem cell to make a variety

of other cells is the focus of stem cell research .

Stem cell therapy, which is still in experimental

stages, involves growing stem cells in the lab

and guiding them toward a desired cell type by

adding different growth factors . The differenti-

ated cells are then surgically implanted . The

theory is that stem cells may then integrate

into the diseased tissue, replace diseased

cells and reverse the effects of the disease .

Cell therapies also could be developed in

which undifferentiated stem cells may be

implanted along with growth factors to guide

their differentiation in the patient’s body . The

aim is to replace the damaged cells with

healthy, disease-free cells—hence the term

regenerative medicine for this approach .

The hope is that stem cells, directed to dif-

ferentiate into specific cell types, could be a

renewable source of replacement cells and

tissues used to treat a wide range of diseases .

Nanotechnology

Nanotechnology deals with the manipulation

of molecules and structures on a nanometer

(one-billionth of a meter) or atomic scale .

Applying nanotechnology for the improvement

of human health is called nanomedicine.

Biotechnology nanomedicine harnesses living

organisms and/or their components on a very

small scale .

One example of nanomedicine is the experi-

mental use of nanoshells to selectively target

and destroy cancer cells at the cellular level .

Nanoshells are nanoscopic metallic lenses

that are selectively delivered to specific

organs or tumors through the bloodstream .

Nanoshells have the ability to capture infrared

light shown through the skin of a cancer patient

and convert it to heat, which kills only the

targeted cancer cells .

Nanoparticles called buckyballs—uniquely

shaped and constructed carbon molecules—

are also showing potential for drug delivery to

target molecules or cells . They may make it

possible to deliver drugs that do not dissolve

in water . Also, because of their small size,

they allow more of the drug to be delivered

per volume . Scientists are working on nano-

particles to unclog blocked arteries .

Future of Biotechnology in Healthcare 33

Future of Biotechnology in Healthcare

New Drug Delivery Systems

Biomedical researchers are studying new

ways of delivering drugs within the body that

could improve effectiveness . One example

is the development of microscopic particles

called microspheres that have tiny holes just

large enough to carry and deliver drugs to

their targets . They are made out of materi-

als that resemble naturally occurring fats in

cell membranes and are delivered as a mist

sprayed into the nose or mouth .

Microsphere therapies are currently available

for lung cancer and respiratory illnesses .

Current research is investigating the use of

microspheres to deliver anticancer drugs to

active tumors and for use with anesthetics

in pain management .

34

The practice of medicine has changed dramatically over the years through pioneering advances in

biotechnology research and innovation, and millions of patients around the globe continue to benefit

from the treatments developed by companies that are discovering, developing and delivering innovative

medicines to treat grievous illnesses. As companies continue to develop medicines that address

significant unmet needs, future innovations in biotechnology research will bring exciting new advances

to help millions more people worldwide.

Looking Ahead

Future of Biotechnology in Healthcare 35

Glossary

Amino Acids: The building blocks of proteins . The unique sequence of amino acids in a chain defines the character of a protein molecule .

Angiogenesis: The process by which the body forms and develops new blood vessels . An-giogenesis can be both beneficial and harmful; while it can be used to stimulate development in new blood vessels to fight clogged arteries, it also allows malignant tumors to increase in size . Angiogenesis is a key area of cancer research .

Antibody: A component of the body’s immune response . A Y-shaped protein, it is secreted in response to an antigenic stimulus . It neutralizes the antigen by binding to it .

Antigen: Any substance, almost always a protein, not normally present in the body that when introduced to the body stimulates a specific immune response and the production of antibodies .

Apoptosis: The process of programmed cell death that may occur in multicellular organisms . Programmed cell death involves a series of biochemical events leading to characteristic cell changes and death . Apoptosis is a key area of cancer research .

Aseptic: Describes a product or method free of microbiological organisms that would lead to contamination .

Assay: A test procedure whereby a property or concentration of a substance is measured .

Autoimmune Disorders: Diseases whereby an individual’s immune system mounts an attack on a portion of its own tissues . Tissues undergoing such an attack can be destroyed in the process .

Base Pairs: Two nucleotides on opposite complementary DNA or RNA strands that are connected via hydrogen bonds . In DNA, adenine (A) forms a base pair with thymine (T), as does guanine (G) with cytosine (C) . In RNA, thymine is replaced by uracil (U) .

Bioinformatics: The application of information technology to the field of molecular biology . Bioinformatics entails the creation and advance-ment of databases, algorithms, computational

and statistical techniques, and theory to solve formal and practical problems arising from the management and analysis of biological data .

Biologic: A product derived from a living organism (from animal products or other biological sources) that is used in the diagnosis, prevention or treatment of disease . Examples of biologics include recombinant proteins, allergy shots, vaccines and hematopoietic growth factors .

Biologic License Application: An application filed with the FDA seeking approval to market a novel biologic in the United States . The application contains a description of the trials and results, formulation, dosage, drug shelf life, manufacturing protocols, packaging information, etc .

Biomarker: A substance used as an indicator of a biologic state . It is a characteristic that is ob-jectively measured and evaluated as an indicator of normal biologic processes, pathogenic pro-cesses or pharmacologic response to a specific therapy . Biomarker identification and measure-ment are regarded as key developments for the future of disease treatment . Biomarkers are also used in drug discovery to determine whether a drug is effective in animal models and at what doses effectiveness is reached .

Biopharmaceutical: A synthetic drug produced utilizing certain biotechnology methods .

Bioreactor: A device or system for growing cells or tissues in the context of cell culture . The pro-cess of fermentation is performed in a bioreactor to grow large volumes of cells for producing specific proteins .

Biosensor: A device that combines a biological component with a physicochemical detector component to detect a pathogenic agent .

Biotechnology: Technology based on biology, especially when used in agriculture, food science and medicine . The United Nations Convention on Biological Diversity defines biotechnology as “any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify products or processes for specific use .”

Glossary

36

Glossary

Blood-Brain Barrier: A physiological mechanism that alters the permeability of brain capillaries so that some substances, such as certain drugs or toxins, are prevented from entering brain tissue, while other substances are allowed to enter freely .

Cell Bank: A facility where cell lines are kept frozen and stored for later use . Cell banks include master cell banks (MCBs) and working cell banks (WCBs) . MCBs house primary cell strains that are kept stored and not used for production purposes . WCBs house cells used in pharmaceutical production grown from those maintained in an MCB so that their stability and uniformity are well characterized .

Cell Culture Technology: The growing of cells outside of living organisms . With mammalian cell culture, it is sometimes possible to replace animal testing with cell testing when evaluating the safety and efficacy of medicines .

Cell Lines: Generations of cells grown from original primary cells . Primary cells are cultured directly from a living organism . With the exception of some derived from tumors, most primary cell cultures have limited life spans . After a certain number of population doublings, cells usually stop dividing, though they remain alive . An established or immortalized cell line has acquired the ability to proliferate indefinitely through either random mutation or deliberate modification .

Cell Viability: Determining whether a cell popula-tion is living or dead . Testing for cell viability usu-ally involves looking at a sample cell population and staining the cells or applying chemicals .

Chemical Library (or Compound Library): A collection of stored chemicals that may be used in high-throughput screening for drug development . The larger the chemical library, the better the chance that high-throughput screen-ing will find a hit (a potential drug candidate) .

Chinese Hamster Ovary Cells (CHO cells): A cell line often used in biological and medical re-search, first introduced in the 1960s . CHO cells are used in studies of genetics, toxicity screen-ing, nutrition and gene expression, particularly expression of recombinant proteins . CHO cells are the most commonly used mammalian hosts for industrial production of protein therapeutics .

Chromatography: A process by which complex

mixtures of different molecules may be sepa-rated from each other . This is accomplished by subjecting the mixture to many repeated partitionings between a flowing phase and a stationary phase .

Chromosome: A threadlike linear strand of DNA and proteins in a cell that houses genes . Chro-mosomes are large enough to be seen under a microscope . In humans, all cells other than germ cells usually contain 46 chromosomes: 22 pairs of autosomes and either a pair of X chromosomes (in females) or an X chromosome and a Y chromosome (in males) . In each pair of chromosomes, one chromosome is inherited from the father and one from the mother .

Clarification: A step in the downstream phase of manufacturing a biologic . After the protein product is harvested, which may include remov-ing intracellular proteins from cells, clarification steps separate the protein from cellular debris . Individual proteins are then separated using chromatography methods .

Clinical Trial: A type of research study that evaluates the safety and efficacy of new drugs, medical devices and biologics in human sub-jects . These tests are required by regulatory agencies as a precondition of regulatory clear-ance to market .

Cloning: The replication of a DNA sequence from one organism to create an exact genetic copy; processes used to create copies of DNA fragments (molecular cloning), cells (cell cloning) or organisms .

Codon: A string of exactly three mRNA bases that code for a specific amino acid during translation of mRNA into DNA .

Colony Hybridization: The screening of a library with a labeled probe (radioactive, biolumines-cent, etc .) to identify a specific sequence of DNA, RNA, enzyme, protein or antibody .

Column Chromatography: A type of chroma-tography that uses a column for containing and separating a mixture . It is a commonly used method of purifying proteins .

Combinatorial Chemistry: A discipline in which a large number of new chemicals are created, compiled into a library and screened for potential therapeutic use .

37

Glossary

Cryopreservation: A process whereby cells or whole tissues are preserved by cooling to low subzero temperatures . At these low tempera-tures, any biological activity, including the bio-chemical reactions that would lead to cell death, is effectively stopped .

DNA (Deoxyribonucleic Acid): DNA is a nucleic acid that contains the genetic information used in the development and functioning of all organ-isms . Molecular systems interpret the sequence of these nucleic acids to produce proteins .

DNA Fingerprinting: A technique used to distinguish between individuals of the same species using only samples of their DNA .

DNA Ligase: The enzyme that creates a bond between the ends of single-stranded DNA segments . Where restriction enzymes are the scissors of recombinant DNA technology, DNA ligase is the glue .

DNA Polymerase: An enzyme that attaches complementary nucleotides to a single stranded human DNA .

DNase (Deoxyribonuclease): Any enzyme that catalyzes the breaking up of linkages in the DNA molecule backbone .

Downstream Phase: Involves manufacturing processes including the recovery, purification, formulation and packaging of the protein .

Enzyme-Linked Immunosorbent Assay (ELISA): A biochemical technique to detect the presence of an antibody or an antigen in a sample . It is commonly used to detect infectious agents .

Enzymes: The many proteins produced by organisms to act as biochemical catalysts . Enzymes are the mediators of cell metabolism .

Epidermal Growth Factor Receptor (EGFR): A cell-surface receptor that is activated when bound by epidermal growth factor . Genetic mutations that lead to EGFR overexpression or overactivity have been associated with a number of cancers .

Extracellular Proteins: Proteins found outside of a cell .

Fermentation: A process of growing, or culturing, cells by using enzymes to effect chemical changes .

Fusion Proteins (or Chimeric Proteins): Proteins created through the joining of two or more genes that were originally coded for separate proteins . Translation of this fusion gene results in a single new protein with functional properties derived from each of the original proteins .

Gel Electrophoresis: A technique used for the separation of DNA, RNA or protein molecules by using an electric current applied to a gel matrix . The gel is the medium used to contain, then separate the target molecules . Electropho-resis refers to the use of electricity to move the molecules through the gel matrix . Placing the molecules in wells in the gel and applying an electric current moves the molecules through the matrix at different rates based on their size, charge and/or shape .

Gene: A length of DNA that codes for a particu-lar protein or, in certain cases, a functional or structural RNA molecule .

Generally Regarded as Safe (GRAS): A designation that a substance is considered safe by experts under the conditions of its intended use . Examples are CHO and NS0 cell lines that have GRAS status for therapeutic protein production .

Genetic Engineering: Alteration of the genetic material of cells or organisms in order, for example, to make them capable of making new substances or performing new functions .

Glycosylation: The process by which oligosac-charide units are added to proteins .

Half-life: A measurement of the time it takes for a drug to lose half of its pharmacologic activity or half of its administered amount in the blood-stream or in its target tissues .

Hematopoietic Growth Factors: Protein hormones produced by the body to regulate blood development, affecting the production and maturation of blood-forming cells .

High-Throughput Screening: The process of screening a sample of compounds rapidly and in parallel, then analyzing the results and choosing further screening compounds based on this information .

Hormones: Substances produced by one tissue and conveyed to another through the blood-stream, usually affecting growth or metabolism .

38

Humanized Antibodies: Monoclonal antibodies that have been synthesized by using recombinant DNA technology to avoid the clinical problem of an immune response to foreign substances . Humanized antibodies are produced by merg-ing the DNA that encodes the binding portion of a monoclonal mouse antibody with human antibody-producing DNA . Cell cultures are used to express this recombinant DNA and produce these partial-mouse and mostly human antibodies .

Hybridization: The process of joining two com-plementary strands of DNA or one each of DNA and RNA to form a double-stranded molecule .

Hybridoma: A cell that has been engineered to produce a desired antibody in large amounts . Hybridomas are created by fusing immortal tu-mor cells with antibody-producing B-lymphocyte cells that continuously synthesize identical (or monoclonal) antibodies .

Immortal Cell Line: An established cell line that has acquired the ability to proliferate indefinitely through either random mutation or deliberate modification .

Immunotherapy: Modulation of the immune system to achieve a therapeutic goal . Monoclonal antibodies are a type of immunotherapy .

Interferon: A naturally occurring cell-signaling protein produced by the immune system in response to infections such as viral infections or parasites .

Intracellular Proteins: Proteins found inside a cell .

Investigational New Drug: A drug that has been approved by the FDA for use in human clinical trials .

In vitro: The technique of performing an experi-ment outside of a living organism, in a controlled environment such as in a cell culture or in cells grown in a petri dish .

Ion Channels: Pore-forming proteins that help establish and control the small voltage gradient across the plasma membrane of all living cells . Ion channels are involved in a wide variety of biological processes and are a favorite target in the search for new drugs .

Mass Spectrometry (MS): An analytic technique for determination of the elemental composition of a sample or molecule . It is also used for de-

termining the chemical structures of molecules, such as peptides or proteins . MS consists of ionizing chemical compounds to generate charged molecules or molecule fragments, then measuring their mass-to-charge ratios .

Media: Nutrient-rich substances in which cells are grown .

Messenger RNA (mRNA): A polynucleotide copy of a DNA gene that communicates the code for building a protein to ribosomes so that new proteins can be built .

Microarray: A tool that enables analysis of the levels of expression of genes in an organism or comparison of gene-expression levels .

Monoclonal Antibody: An antibody produced by cells that are all derived from a single antibody-producing cell . Once a cell capable of generating an antibody with desired therapeutic characteristics is selected, laboratory processes are used to clone (make large numbers of) these cells . Since the cells are all identical and are produced by cloning one specific cell in great numbers, they are called monoclonal and can be used to continuously produce identical antibody molecules with these same therapeutic characteristics .

Nanomedicine: The medical application of nanotechnology .

Nanotechnology: The study and creation of systems and devices at the level of molecules and atoms .

Neurotransmitters: Chemicals that are used to relay, amplify and modulate signals between a neuron and another cell .

Non Secreting (NS0) Cells: Mouse myeloma cells that are used frequently in the production of recombinant antibodies .

Nucleotide: The name given to an individual unit of the DNA double helix and RNA . A nucleotide contains one sugar, one phosphate and one base .

Nucleus: The organelle within a living cell that contains genetic material and controls life functions .

Pathogen: A disease-causing agent such as a bacterium or virus .

Pegylation: The process of adding polyethylene glycol to a therapeutic protein, which enables

Glossary 39

Glossary

the therapeutic protein to stay in the body longer .

Peptibodies: Engineered therapeutic fusion proteins with attributes of both peptides and antibodies but that are distinct from each and that can bind to human drug targets .

Peptide Bond: A bond that links together two or more amino acids . A protein is a long chain of amino acids joined together with peptide bonds and therefore is sometimes referred to as a polypeptide .

Peptides: Short chains of amino acids . Poly-peptides, or multiple peptides linked together by peptide bonds, are long chains of amino acids .

Personalized Medicine: Use of the information contained within a patient’s genome, genotype or genomic signature to design and tailor the best treatment plan for that individual patient .

Pharmacodynamics: Studies performed to determine what a drug does to the body .

Pharmacogenomics: The science of under-standing the correlation between patients’ genetic makeup (genotype) and their responses to drug treatment .

Pharmacokinetics: Studies performed to deter-mine what the body does to a drug .

Phosphorylation: The addition of a phosphate (PO4 ) group to a protein or other organic mol-ecule . Protein phosphorylation plays a significant role in a wide range of cellular processes .

Polymerase Chain Reaction (PCR): A method for creating millions of copies of a particular seg-ment of DNA . If a scientist needs to detect the presence of a very small amount of a particular DNA sequence, PCR can be used to amplify the amount of that sequence until there are enough copies available to be detected .

Preclinical Trials (or Studies): Tests that take place in a scientifically controlled setting using cell culture and/or animals as disease models .

Product Pipeline: In the biomedical industry, the term pipeline refers to the number of unique products or processes reported or in develop-ment by a company . Drugs that have entered into clinical trials are said to be “in the pipeline .”

Protein Engineering: A process for isolating and studying proteins and generating proteins with

modified structures by altering the genes that direct their composition .

Proteins: Compounds (chains of amino acids) constituting the ultimate expression product of a gene . Created through the synthesis performed by ribosomes, proteins are the workhorses of living systems, causing chemical processes and changing as their environment changes .

Proteomics: The study of proteins . Proteomics has three major goals: to identify and quantify all the proteins expressed in an organism, to determine the structure and function of each protein and to study the protein-protein interac-tions that affects how one protein interacts with other proteins to control cellular processes .

Receptor (Cell Receptor): A protein molecule, embedded in either the plasma membrane or the cytoplasm of a cell, to which a mobile signaling (or signal) molecule may attach . A molecule that binds to a receptor is called a ligand, and may be a peptide (such as a neurotransmitter), a hormone, a pharmaceutical drug or a toxin, and when such binding occurs, the receptor goes into a conformational change, which usually initiates a cellular response .

Recombinant DNA: A form of DNA that does not exist naturally and is created by combining DNA sequences that would not normally occur together .

Recombinant Proteins: Proteins created by recombinant DNA technology .

Regenerative Medicine: Research into treatments that restore damaged cells with healthy, disease-free cells .

Restriction Enzymes: Enzymes having the ability to cut DNA at a certain location . Scientists use these enzymes to isolate certain types of DNA and place them into new environments . Where DNA ligase is the glue of recombinant DNA tech-nology, restriction enzymes are the scissors .

Reverse Transcriptase: An enzyme used by retroviruses to form a complementary DNA se-quence (cDNA) from an RNA template—usually the genome of the retrovirus . The enzyme then performs a complementary template of the cDNA strand such that a double-stranded DNA molecule is formed . This double-stranded DNA molecule is then inserted into the chromosome

40

of the host cell, which has been infected by the retrovirus .

Ribonucleic Acid (RNA): A molecule similar to DNA, which helps in the process of decoding the genetic information carried by DNA . RNA is a nucleic acid transcribed from DNA; mRNA is then translated into proteins .

Ribosome: The cell structures within which protein synthesis occurs .

RNA Interference: A mechanism that inhibits gene expression at the stage of translation (see translation) or by hindering the transcription (see transcription) of specific genes . This method has been referred to as posttranscrip-tional gene silencing and is an important tool for gene-expression research .

Signal Transduction: The movement of signals from the outside of a cell to the inside . Scientists are attempting to learn more about this process in cancer cells in order to fight the disease .

Single-Nucleotide Polymorphism (SNP): A DNA sequence variation that occurs when a single nucleotide—A, T, G or C—in the genome dif-fers between members of a species . Variations in the DNA sequences of humans can affect how humans develop diseases and respond to pathogens, chemicals, drugs, vaccines and other agents .

Southern Blotting: Transfer by absorption of DNA fragments separated in electrophoretic gels to membrane filters for detection of specific base sequences by radiolabeled complementary probes .

Stem Cell: Undifferentiated, human cells with the ability both to multiply and to differentiate into specific cells .

Thermocycler: A laboratory apparatus, used to amplify segments of DNA via the polymerase chain reaction (PCR) process . The device has a thermal block with holes where tubes holding the PCR mixtures can be inserted . The cycler then raises and lowers the temperature of the block in discrete, preprogrammed steps .

Transcription: The process by which enzymes use the genetic information on a strand of DNA to create a complementary strand of messenger RNA .

Transfer RNA: Molecules that carry amino acids during the process of protein synthesis during translation .

Transformation: The process of transferring DNA from a donor to a recipient cell . Scientists use this process to introduce recombinant DNA to bacteria, yeast and mammalian cell lines .

Transgenic: A term describing an organism containing genetic material from a source other than its parents .

Translation: The process that converts an mRNA sequence into a string of amino acids that form a protein . Translation follows transcription (see transcription).

Upstream Phase: Involves the production of the protein product, most often by using cells (mi-crobial, insect or mammalian) growing in culture .

Vaccine: An agent bearing antigens on its surface that causes activation of the immune system without causing actual disease .

Vector: (1) An organism that serves to transfer a disease-causing organism (pathogen) from one organism to another . (2) A mechanism whereby foreign genes are moved into an organism and inserted into that organism’s genome .

X-ray Crystallography: A method of determin-ing the arrangement of atoms within a crystal, in which a beam of X-rays strikes a crystal and scatters in many different directions . From the angles and intensities of these scattered beams, a crystallographer can produce a three-dimen-sional picture of the density of electrons within the crystal, and the structure of a substance can be determined .

Glossary 41

Timeline of Medical Biotechnology

1950s1952

- Dr . George Gey establishes a continuous cell line taken from a human cervical carcinoma isolated from Henrietta Lacks, who died of the cancer in 1951 . This cell line, containing HeLa cells, is commonly used in medical research .

1953

- Dr . James Watson and Dr . Francis Crick reveal the 3-D structure of DNA .

- Dr . Joseph Murray performs the first kidney transplant between identical twins .

1955

- An enzyme, DNA polymerase, involved in the synthesis of a nucleic acid, is isolated for the first time .

- Dr . Jonas Salk develops the first polio vaccine . The development marks the first use of mam-malian cells (monkey kidney cells) and the first application of cell culture technology to generate a commercial product .

1957

- Scientists prove that sickle-cell anemia occurs due to a change in a single amino acid .

1958

- Dr . Arthur Kornberg of Washington University in St . Louis makes DNA in a test tube for the first time .

- The first automatic protein sequencer, the Moore-Stein amino acid analyzer, is developed .

1960s1960

- French scientists discover messenger RNA (mRNA) .

1961

- Scientists understand genetic code for the first time .

1962

- Dr . Osamu Shimomura of Boston University discovers the green fluorescent protein Aequorea victoria in jellyfish . He later develops

it into a tool for observing previously invisible cellular processes .

1963

- Independent groups in the United States, Germany and China synthesize insulin, a pancreatic hormone .

1964 - The existence of reverse transcriptase is

predicted .- Dr . Samuel Katz and Dr . John F . Enders

develop the first vaccine for measles .

1967

- Dr . Maurice Hilleman develops the first American vaccine for mumps .

1969

- The first vaccine for rubella is developed . It is combined with the measles and mumps vaccines to form the measles/mumps/rubella vaccine in 1972 .

1970s1970 - Restriction enzymes are discovered . These

enzymes cut DNA into pieces and are used for various studies and applications . The restriction enzyme technique becomes a fundamental tool in modern genetic research and opens the way for gene cloning.

- Dr . Har Gobind Khorana synthesizes the first complete gene at the University of Wisconsin–Madison .

1972

- DNA ligase, which links DNA fragments together, is used for the first time .

- The DNA composition of humans is discovered to be 99 percent similar to that of chimpanzees and gorillas .

- The purified enzyme reverse transcriptase is first used to synthesize complementary DNA from purified messenger RNA in a test tube .

1973 - Dr . Stanley Cohen and Dr . Herbert Boyer use

bacterial genes to perform the first successful recombinant DNA experiment, which inserted

Timeline of Medical Biotechnology

42

Timeline of Medical Biotechnology

a recombinant DNA molecule into a cell for replication .

- Dr . Edwin Southern develops a blotting technique for DNA called the Southern blot . It becomes a seminal technology for studying the structure of DNA .

1974 - The U .S . National Institutes of Health (NIH)

forms a Recombinant DNA Advisory Commit-tee to oversee recombinant genetic research .

- The first vaccine for chicken pox is developed in Japan .

1975

- Colony hybridization and Southern blotting are developed for detecting specific DNA sequences .

- The first monoclonal antibodies are produced . Dr . César Milstein, Dr . Georges Kohler and Dr . Niels Jeme develop monoclonal antibody technology by fusing immortal tumor cells with antibody-producing B-lymphocyte cells to pro-duce hybridomas that continuously synthesize identical (or monoclonal) antibodies .

1976

- The NIH releases the first guidelines for recom-binant DNA research .

- Molecular hybridization is used for the prenatal diagnosis of alpha thalassemia .

- Yeast genes are expressed in E. coli bacteria .

1977 - Protocols are developed to rapidly sequence

long sections of DNA .- Genetically engineered bacteria are used

to synthesize the human growth protein somatostatin, marking the first time a synthetic recombinant gene is used to clone a protein . Many consider this to be the advent of the Age of Biotechnology .

- Dr . Robert Austrian of the University of Pennsylvania develops the first vaccine for pneumonia .

1978

- Dr . Herbert Boyer of the University of California, San Francisco, constructs a synthetic version of the human insulin gene and inserts it into the bacterium E. coli, allowing the bacterium to produce human insulin .

- The first test-tube baby, Louise Brown, is born in the United Kingdom .

- The first vaccine for meningococcal meningitis is developed .

1980s1980

- The U .S . Supreme Court rules genetically altered life forms can be patented, opening up enormous possibilities for commercially exploit-ing genetic engineering . The first patent of this nature was awarded to the Exxon oil company to patent an oil-eating microorganism, which would later be used in the 1989 cleanup of the Exxon oil spill at Prince William Sound, Alaska .

- Dr . Stanley Cohen and Dr . Herbert Boyer receive a U .S . patent for gene cloning .

- The first automatic gene machine, or gene- synthesizing machine, is developed in California .

- Founding of Amgen, which will grow to become the world’s largest biotechnology medicines company .

1981

- Dr . Baruch Blumberg and Dr . Irving Millman develop the first vaccine for hepatitis B (not recombinant) four years after the virus is discovered .

- Scientists in Switzerland clone mice . - The first transgenic animals are produced

by transferring genes from other animals into mice .

1982 - The U .S . Food and Drug Administration (FDA)

approves the first biologic—or recombinant protein: Humulin®, Genentech’s human insulin drug produced by genetically engineered bac-teria for the treatment of diabetes .

1983 - Dr . Luc Montagnier of the Pasteur Institute in

Paris isolates the AIDS virus .- Dr . Kary Banks Mullis invents the polymerase

chain reaction (PCR), a technique for multiply-ing DNA sequences . PCR is recognized as the most revolutionary molecular biology technique of the 1980s .

- The FDA approves a monoclonal antibody-based diagnostic test to detect Chlamydia trachomatis.

- The first artificial chromosome is synthesized .- The first genetic markers for specific inherited

diseases are found .

43

Timeline of Medical Biotechnology

1984 - The DNA fingerprinting technique is devel-

oped . When a restrictive enzyme is applied to DNA from different individuals, the resulting sets of fragments sometimes differ markedly from one person to the next . Such variations in DNA are called restriction fragment length polymorphisms and are extremely useful in genetic studies .

- The first genetically engineered vaccine is developed for hepatitis B .

- The entire genome of the HIV virus is cloned and sequenced .

1985 - Genetic fingerprinting enters the courtroom .- Genentech becomes the first biotechnology

company to launch its own biopharmaceutical product .

- Genetically engineered plants resistant to insects, viruses and bacteria are field-tested for the first time .

- Cloning of the gene that encodes human lung surfactant protein is accomplished . This is a major step toward reducing premature birth complications .

- The NIH approves guidelines for performing experiments in gene therapy on humans .

1986 - University of California, Berkeley, chemist

Dr . Peter Schultz describes how to combine antibodies and enzymes (abzymes) to create therapeutics .

- The automated DNA sequencer is invented in California .

- The FDA approves the first monoclonal antibody treatment to fight kidney transplant rejection .

- The FDA approves first biotech-derived interferon drugs to treat cancer . In 1988, the drugs are used to treat Kaposi’s sarcoma, a complication of AIDS .

- The FDA approves the first genetically engineered human vaccine to prevent hepatitis B .

1987 - The FDA approves a genetically engineered

tissue plasminogen activator to treat heart attacks .

- Dr . Maynard Olson and colleagues at Washington University invent yeast artificial

chromosomes, which are expression vectors for large proteins .

- Reverse transcription and the polymerase chain reaction are combined to amplify messenger RNA sequences .

- DNA microarray technology, the use of a collection of distinct DNAs in arrays for expression profiling, is first described . The arrayed DNAs are used to identify genes whose expression is modulated by interferon .

- The FDA approves a diagnostic serum tumor marker test for ovarian cancer .

1988

- Congress funds the Human Genome Project, a massive effort to map and sequence the human genetic code as well as the genomes of other species .

- The first agreement between two companies with parallel patents for cross-licensing of biotech products occurs and becomes the prototype .

1989 - The FDA approves Amgen’s first biologically

derived human therapeutic .- Oil-eating bacteria are used to clean up the

Exxon Valdez oil spill .- A gene responsible for cystic fibrosis is

discovered .

1990s1990

- The first federally approved gene therapy treat-ment is performed successfully on a 4-year-old girl suffering from an immune disorder called adenosine deaminase deficiency .

- The Human Genome Project, the international effort to map all of the genes in the human body, is launched . Estimated cost: $13 billion .

- The FDA licenses the first hepatitis C antibody test, which helps to ensure the purity of blood bank products .

- The FDA approves a bioengineered form of the protein interferon gamma to treat chronic granulomatous disease .

- The FDA approves a modified enzyme for enzyme replacement therapy to treat severe combined immunodeficiency disease . It is the first successful application of enzyme replace-ment therapy for an inherited disease .

44

1992

- The U .S . Army collects blood and tissue sam-ples from all new recruits as part of a genetic dog tag program aimed at better identification of soldiers killed in combat .

- The FDA approve the first genetically engineered blood-clotting factor—a recombinant protein used to treat hemophilia A .

- The FDA approves a recombinant protein to treat renal cell cancer .

- American and British scientists unveil a tech-nique for testing embryos in vitro for genetic abnormalities such as cystic fibrosis and hemophilia .

1993 - The FDA approves a recombinant protein to

treat multiple sclerosis—marking the first new multiple sclerosis treatment in 20 years .

- An international research team, led by Dr . Daniel Cohen of the Center for the Study of Human Polymorphisms in Paris, produces a rough map of all 23 pairs of human chromosomes .

- Two smaller trade associations merge to form the Biotechnology Industry Organization, an international biotechnology advocacy group .

1994

- The FDA approves a recombinant protein to treat growth hormone deficiency .

- Dr . Mary-Claire King at the University of California, Berkeley, discovers the first breast cancer gene, BRCA1 .

- The FDA approves a modified enzyme to treat Gaucher’s disease .

- A multitude of genes, human and otherwise, are identified and their functions described . These include:

•Ob,agenepredisposingtoobesity •BCR,abreastcancersusceptibilitygene •BCL-2,ageneassociatedwithapoptosis

(programmed cell death) •Hedgehoggenes(namedbecauseoftheir

shape) produce proteins that guide cell differentiation in advanced organisms

•Vpr,agenegoverningreproductionoftheHIV virus

- Linkage studies identify genes for a variety of ailments, including bipolar disorder, cerulean cataracts, melanoma, hearing loss, dyslexia, thyroid cancer, sudden infant death syndrome, prostate cancer and dwarfism .

- The FDA approves a genetically engineered

version of human DNase, which breaks down protein accumulation in the lungs of cystic fibrosis patients . It represents the first new therapeutic drug for managing cystic fibrosis in more than 30 years .

1995

- The first baboon-to-human bone marrow transplant is performed on an AIDS patient .

- The first vaccine for hepatitis A is developed . The NIH, the U .S . Army and the Centers for Disease Control and Prevention are significantly involved in the development and clinical testing of the vaccine .

- Scientists at the Institute for Genomic Research complete the first full gene sequence of a living organism (other than a virus) for the bacterium Haemophilus influenzae.

- A European research team identifies a genetic defect that appears to be the most common cause of deafness .

1996

- The Department of Biochemistry at Stanford University and Affymetrix develop the GeneChip, a small glass or silica microchip that contains thousands of individual genes that can be analyzed simultaneously . This marks a techno-logical breakthrough in gene expression and DNA-sequencing technology .

- A group of scientists sequence the complete genome of a complex organism, Saccharomyces cerevisiae, otherwise known as baker’s yeast . The achievement marks the complete sequencing of the largest genome to date—more than 12 million base pairs of DNA .

- A new, inexpensive diagnostic biosensor test is developed to allow instantaneous detection of a toxic strain of E. coli, the bacteria responsible for many food-poisoning outbreaks .

1997 - The first human artificial chromosome is cre-

ated . A combination of natural and synthetic DNA is used to create a genetic cassette that can potentially be customized and used in gene therapy .

- The FDA approves a recombinant follicle- stimulating hormone to treat infertility .

- The FDA approves the first bloodless HIV-anti-body test that uses cells from patients’ gums .

- Scientists at the Institute for Genomic Research sequence the complete genome of the Lyme

Timeline of Medical Biotechnology 45

Timeline of Medical Biotechnology

disease pathogen, Borrelia burgdorferi, along with the genome for the organism linked to stomach ulcers, Helicobacter pylori.

- Scientists at the University of Wisconsin– Madison sequence the E. coli genome .

- The FDA approves the first therapeutic antibody to treat cancer in the United States . It is used for patients with non-Hodgkin’s lymphoma .

1998 - Human skin is produced for the first time in

the lab .- Two research teams culture embryonic stem

cells . Embryonic stem cells are used to regen-erate tissue and create disorders mimicking diseases .

- Scientists at the Sanger Institute of the Washington University School of Medicine in St . Louis sequence the first complete animal genome for the Caenorhabditis elegans worm .

- A rough draft of the human genome map is produced, showing the locations of more than 30,000 genes .

- The first vaccine for Lyme disease is developed .- The FDA approves a novel monoclonal anti-

body to treat Crohn’s disease .- A monoclonal antibody therapy used against

breast cancer has favorable results, heralding a new era of treatment based on molecular targeting of tumor cells .

- Approval of the Her-2 inhibitor for the treatment of breast cancer patients who have tested positive for the Her-2 mutation brings personalized medicine to oncology .

1999 - The complete genetic code of the human

chromosome is deciphered .

2000s2000 - Scientists at Celera Genomics and the Human

Genome Project complete a rough draft of the human genome .

2001

- Science and Nature magazines publish the hu-man genome sequence, making it possible for scientists all over the world to begin research-ing new treatments for diseases that have genetic origins, such as cancer, heart disease, Parkinson’s and Alzheimer’s .

2002 - An era of very rapid shotgun sequencing of

major genomes is completed . Included are the mouse, chimpanzee, dog and hundreds of other species .

2003 - Celera and NIH complete sequencing of the

human genome .

2004 - The FDA approves the first monoclonal anti-

body that is an antiangiogenic, inhibiting the growth of blood vessels—or angiogenesis—for cancer therapy .

- The FDA clears a DNA microarray test system, which aids in selecting medications for a variety of conditions . This is an important step toward personalized medicine .

2006 - The FDA approves a recombinant vaccine

against human papillomavirus, which causes genital warts and can cause cervical cancer .

- Scientists determine the 3-D structure of the human immunodeficiency virus, which causes AIDS .

2007 - Scientists discover how to use human skin

cells to create embryonic stem cells .

2008 - Chemists in Japan create the first DNA mol-

ecule made almost entirely of artificial parts . The discovery can be used in the fields of gene therapy .

- Dr . Craig Venter and his team replicate a bacterium’s genetic structure entirely from laboratory chemicals, moving a step closer toward creating the world’s first living artificial organism .

46

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