Biodefense Vaccines, Therapeutics and

Diagnostics

NGIC Workshop

September 27, 2004

Robert L. McKown, Ph.D.

What is Biotechnology?

What is Biotechnology?

Recombinant genetic Engineering…using biological process to develop products

G. Steven Burrill 1997

GeneticEngineering

RawMaterials

LivingSystem

Product

Enabling Technology

Recombinant DNA TechnologyBoyer and Cohen, 1973

The Birth of an Industry

University of California, San Francisco

Genentech Inc. South San Francisco

Founded April 7, 1976Herbert Boyer and Robert Swanson

Proof of Concept

• Cloning the human insulin gene• Cloned by Genentech scientists in 1978• Licensed to Eli Lilly• First recombinant drug marketed, 1982• Product revenues from human insulin

(humulin) reached $900 million in 2003

The Promise of Biotechnology

Bioproduction of molecules so complex they can only be synthesized in a living system

DNA PROTEIN

Biotech Products

The JMU Biomanufacturing Laboratory

Concept document created in 1997

Development funding awarded in 1998

Lab space committed in 1999

Lab design and set up in 2000

Became operational in 2001

Founding Sponsors

Biomanufacturing Technology

Genetic manipulations and analysis

Cell line development

Fermentation and cell culture technology

Product purification technology

Quality assurance and quality control

Regulatory guidelines and compliance

Process development

The Production Platform

Cell LineDevelopment

Systems ManagementRegulatory AffairsQuality Assurance

Production

Product Recovery

Product Analysis

Quality ControlAnalytical Testing

Documentation

                              

 

Development of Expression Systems

Cell Line Development

Production and Purification

Product Analysis

Proof of Concept

Cloning, Expression, & Purificationof Green Fluorescence Protein (GFP)

Purification of GFP

GFP Purification: Sephadex G-75Chromatography

0

0.1

0.2

0.3

0.4

0.5

0.6

1 4 7 10 13 16 19 22 25 28 31 34 37 40Fraction Number

Ab

sorb

an

ce 2

80 n

m

0

0.02

0.04

0.06

0.08

0.1

Ab

sorb

an

ce 3

95 n

m

A280

A395

Analysis of GFP

Fraction I II III IV STD

Mol. Wt. 94,000 67,00043,000

30,000 20,100 14,400

                       

 

Projects and Collaborations

• Lacritin project, UVA and EyeRx

• Vaccine and diagnostics projects, USAMRIID

• Staphylococcus aureus Protein A project, Genentech, Inc.

• New River Pharmaceuticals project

• M. tuberculosis project, Veterans Hospital, Long Beach, CA

Research and Development of a Recombinant Protein

(Lacritin) as a New Therapeutic for Treatment

for Dry Eye Syndrome

Lacritin and the Lacrimal Gland

A stable, novel human tear glycoprotein

Produced primarily by the lacrimal gland located on the upper eyelid

Also found in salivary glands and the thyroid

Properties of Lacritin Demonstrated in Cell Culture

Lacritin hydrates the corneal surface as a natural component of tears

Lacritin causes the rapid, sustained release of calcium in cultured corneal epithelial cells

Lacritin stimulates proliferation of ductal cells

Potential as a Therapeutic Agent

• Dry Eye Syndrome• Caused by the decrease in both the quality and

volume of tear secretion

• Affects over 35 million Americans

• The application of eye drops is the primary current treatment

• Lacritin as a Potential Therapeutic• Promotion of tear production

• Induction of cell proliferation

Research History

• Lacritin gene discovered and characterized by the Gordon Laurie lab at UVA

• JMU collaboration with the Laurie lab

• Engineered a recombinant mature lacritin gene that was cloned, expressed, and purified

• Current Research

• Creation of C-terminal deletions to identify domains of biological activity

• Lacritin protein in pre-clinical animal studies for treatment of Dry Eye Syndrome

Future Applications of Biotechnology

• New Human Therapeutics and Vaccines• New Human Diagnostics• New Genetically Engineered Microbes,

Plants, and Animals• New Biomaterials (Rubber, Plastics,

Spider Silk)• Genetically Engineered Humans?

Acknowledgements

• Robert McKown, Ph.D., JMU

• Ron Raab, Ph.D., JMU

• George Coffman, Ph.D., JMU

• Gordon Laurie, Ph.D., UVA

• Jeremy Goodin, Ph.D., USAMRIID

• Patricia Williams, Ph.D., EyeRx

• JMU Students