Immunogenicity TestingTechnologies and Services

CONSISTENT REPRODuCIBLE

RESuLTS

BIOLOGICSEXPERTISE

AlphaLISA® . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

DELFIA® . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

ELAST® . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

Detection and Automation Solutions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

OnPointSM Reagent Services . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

Ordering Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

Table of Contents

www.perkinelmer.com/immunogenicity

SENSITIVETO THE NEED FOR

RELIABLETESTING

1

To ensure researchers have the right resources, PerkinElmer offers an immunogenicity testing portfolio designed to provide a complete solution, from multiple assay platforms to automation and detection instruments and custom assay development. Our distinct yet complementary assay platforms include no-wash, bead-based AlphaLISA, DELFIA time-resolved fluorescence (TRF) immunoassays and ELAST ELISA signal amplification.

Plate to plate, assay to assay, operator to operator, PerkinElmer gives you the tools you need to ensure the consistent, reproducible results your research requires and patient safety demands.

Immunogenicity has critical implications for the safety and efficacy of the therapeutic proteins increasingly used in drug development. Potential patient immune responses

to these biopharmaceuticals vary widely from little effect to life-threatening. So the development of valid, sensitive assays to detect anti-drug antibodies is vital. Commonly used technologies for immunogenicity testing include standard ELISA and electrochemiluminescence; however, each has shown limitations when it comes to accurate and consistent detection of anti-drug antibodies.

Log [TNFa(g/mL)]

DELFIA

Range, Sensitivity, Stability

1000000

100000

10000

1000

-13 -12 -11 -10 -9 -8 -7100

ELASTSee What You’ve Been Missing in Your ELISA

AlphaLISA

The No-wash ELISA

Standard ELISA

Think Outside the Well

Scal

ed u

nits

PerkinElmer technologies provide greater sensitivity and broader dynamic range than ELISA .

ELISA-related Technologies

2

NO-WASHALPHALISATHE BETTER WAYTO ASSAY

AlphaLISA is a homogeneous alternative to conventional ELISA, enabling you to eliminate tedious wash steps without sacrificing high sensitivity or wide dynamic range. Part of PerkinElmer’s

exclusive Alpha Technology platform, it is a bead-based, proximity assay that does not require washing or separation.

When biological interactions bring Alpha Donor and Acceptor beads into close proximity, a greatly amplified signal is produced, which is directly proportional to the amount of anti-drug antibody or biomarker present in the sample. The beads are extremely versatile and can be coated with a wide variety of biomolecules, allowing the highly sensitive detection of unique biological events.

Emission615 nm

Excitation680 nm

Streptavidin-coatedAlpha Donor Bead

Drug IgG-coatedAlphaLISA Acceptor Bead

BiotinylatedDrug IgG

Anti-DrugIgG (ADA)

1. The Donor and Acceptor beads can be coated with different molecules depending on the type of reaction or event that needs to be studied.

2. Interaction between bead-bound molecules and analyte brings Donor and Acceptor beads into close proximity.

3. Donor bead excitation (680 nm) generates release of up to 60,000 singlet oxygen molecules per bead per second, a signal amplification that produces the high sensitivity intrinsic to Alpha Technology.

4. The singlet oxygen molecules released from the Donor beads react with the Acceptor beads to produce a luminescent emission (615 nm) that is proportional to the amount of analyte present in solution.

5. Because singlet oxygen can travel up to 200 nm in solution, Alpha Technology can be used to study interactions between very large biological molecules.

Mechanism of Action

The US Food and Drug Administration currently recommendsthat screening assays achieve a sensitivity of approximately250-500 ng/mL . This concentration corresponds to the highend of the AlphaLISA calibration curve .

3www.perkinelmer.com/immunogenicity

AlphaLISA: The fastest, easiest assays ever

• Homogeneous: No wash steps are required, so low-affinity anti-drug antibodies can be detected which would be washed away in a heterogeneous assay format.

• High sensitivity: The signal is greatly amplified, allowing very small volumes of sample to be used (1-5 µL).

• Wide dynamic range: Reduces the need for dilution steps.

• Good drug tolerance: The presence of free drug in the sample has minimal impact on the detection of the anti-drug antibody.

• High throughput: Allows a higher density microplate format to be used, increasing the number of samples per plate, and reducing the number of plates and cutoff point determinations.

• Automatable: Reduces risk of error and increases reproducibility.

• Cost-effective: Small amounts of sample and reagents required.

Choose Your Donor Bead

Choose Your Acceptor Bead

Add Your Antibodies Go!

Streptavidin Protein A Protein A Nickel Chelate

Glutathione Unconjugated Protein G Glutathione (GSH)

Nickel Chelate Anti-human IgG (Fc-specific) Anti-GST

Unconjugated Anti-rabbit IgG (Fc-specific) Anti-c-myc

Anti-mouse IgG (Fc-specific) Anti-FLAG

Anti-rat IgG (Fc-specific) Anti-DIG

Anti-goat IgG (Fc-specific) Unconjugated

Alpha Donor Beads AlphaScreen® Acceptor Beads* AlphaLISA Acceptor Beads

Alpha Technology Toolbox Beads for Immunogenicity Testing

How to use the Alpha Technology Toolbox

Custom coatings and conjugations are available . Please see OnPoint Reagent Services on page 7 for more detail .

*AlphaScreen is another Acceptor bead type, which emits at 520-620 nm .

4

DELFIA: The right selection for sensitiveresearch assays

• Very high sensitivity: Allows detection of low concentrations of anti-drug antibodies; less sample and reagent required.

• Wide dynamic range: No need for additional dilutions.

• High signal stability: Improves reliability and ease of use.

• Easy to convert: Similar format to ELISA, allowing easy conversion.

• Cost-effective: High sensitivity and familiar workflow make switching to DELFIA easy.

DELFIARAISE YOuREXPECTATIONS

When it comes to range, sensitivity and stability, DELFIA is the optimal choice. It is a proven, robust immunodetection platform

based on time-resolved fluorescence (TRF) detection. This detection method, coupled with the unique chemical properties of lanthanide chelates, allows DELFIA to deliver higher sensitivity, a broader dynamic range, good drug tolerance and superior stability compared to standard ELISA. Furthermore, converting your ELISA-based immunogenicity assay could not be easier, as the assay designs for ELISA and DELFIA are virtually identical. DELFIA is complementary to Alpha Technology, and is the ideal choice when a highly sensitive, cost-effective assay format is required.

1. The DELFIA assay principle is virtually identical to that of a standard sandwich ELISA; analyte is first captured on a coated microplate, followed by addition of detection antibody to complete the sandwich.

2. Unlike ELISA, DELFIA utilizes a lanthanide chelate-labeled detection antibody, which exhibits minimal fluorescence by itself.

3. An enhancement step unique to DELFIA releases the lanthanide from the antibody complex, producing a new, highly fluorescent lanthanide chelate contained within a protective micelle.

4. The amplified fluorescent signal is detected using time-resolved fluorometry. This detection method removes non-specific interfering fluorescent background signal and provides DELFIA the superior sensitivity it is known for.

Mechanism of Action

Eu

Eu

340 nm

5www.perkinelmer.com/immunogenicity

ELASTAMPLIFYYOuR ELISA

See what you’ve been missing from your ELISA assays. ELAST (ELISA Amplification System) is designed to amplify the signal generated by the enzyme horseradish peroxidase (HRP) in solid

phase analytical methods such as ELISA. ELAST is based on PerkinElmer’s proprietary TSA™ Tyramide Signal Amplification system. Any ELISA assay using HRP can be enhanced using ELAST, returning an 8- to 32-fold signal increase. Requiring only an additional 30 minutes to the ELISA protocol, ELAST can dramatically reduce the consumption of precious antibodies and samples.

ELAST: Save resources while increasing ELISA signals

• High sensitivity: Amplify your ELISA signal 8- to 32-fold.

• Convenient: Easily integrates into your existing ELISA protocol.

• Cost-effective: Minimize antibody and sample use.

• Proven: Powered by PerkinElmer’s proprietary TSA technology.

PerkinElmer technologies take you beyond ELISA for better results in immunogenicity.

More stars are better .

Sensitivity Throughput Detection of Low- affinity Abs

Sample Consumption

Drug Tolerance

Matrix Interference Tolerance

Bridging ELISA * * * * * *

AlphaLISA *** *** *** *** *** ***

DELFIA *** ** ** ** *** ***

ELAST ELISA ** * * ** * *

HRP

T

HRP

T

HRP HRP

HRP

T

HRP

T

HRP

T

HRP

T

HRP T

HRP T

HRP T

HRP T

HRP

T

HRPT

HRP

T

HRP

T

HRP

T

HRP

T

StandardELISA

ELASTELISA

HRP

T

HRP

T

6

DETECTION ANDAuTOMATIONCOME TOGETHER FORREPRODuCIBLE

RESuLTSWhy trust your critical experiments, and precious samples and antibodies to anyone else? PerkinElmer develops all its assay platforms on our detection instrumentation, so you can be confident that you will get consistent results, every time. And we’re the exclusive provider of integrated, validated Alpha Technology solutions.

EnVision®

EnSpire™

Multilabel Plate Readers Microplates

PerkinElmer is the company to go to first for all yourmultilabel detection needs . With plate readers that fit every application and budget, PerkinElmer’s expertise ensures performance and value for optimal investigating, screening and discovery .

Automated Liquid Handling

Easy automation: PerkinElmer’s liquid handling instruments deliver ease of use and flexibility to complement a wide array of application techniques to support your requirements in precision dispensing, microplate capacity and dynamic volume range .

Automation and Detection Solutions to Enhance Your Assay Performance

Seamless integration: Prepare your assays with the JANUS, and select the EnSpire or EnVision plate reader for more efficient sample preparation and reproducible results . Plate to plate, assay to assay, operator to operator, with PerkinElmer you can always count on our reliable results .

PerkinElmer microplates are designed specifically for use with our own assays and detection instruments . We validate all of our assays on PerkinElmer microplates .

JANUS® Automated Workstation

7www.perkinelmer.com/immunogenicity

ONPOINTREAGENT SERVICESWE DEVELOPYOu DISCOVER

We understand that no two immunogenicity projects are exactly the same. Whatever your requirements, we can offer proof-of-concept or design a customized solution

for you. OnPoint Services scientists are experts in PerkinElmer’s state-of-the-art assay technologies and detection instrumentation. They know our products and equipment inside and out and pride themselves in applying their technical knowledge to help you reach your critical research goals.

The OnPoint team is able to recommend solutions from one of the industry’s broadest portfolios of reliable and innovative technologies, the choice of many leading biotechnology and pharmaceutical companies worldwide.

On Pointservices

FPOOnPoint Services

On Pointservices

Validated Microplates

EnVision Multilabel

Plate Readers

EnSpire Multilabel Plate Readers

JANUS Automated

Workstations

Sensitive, Reliable Assays

From assays to automation, only PerkinElmer offers a validated, complete solution for immunogenicity testing .

OnPoint’s range of services includes:

• Assay development and proof-of-concept for immunogenicity assays

• Custom labeling services• Custom microplate barcoding and coating

Because we focus on your success and have the ability to choose from such a wide range of assay technologies, we custom design the right solution for you – the one that meets your specific assay requirements.

Find out more at www.perkinelmer.com/onpoint.

Your Complete Immunogenicity Solution

8

ORDERING INFORMATION

Alpha ToolboxProduct Information Product Description Size Part Number

Alpha Donor Beads(order Alpha Acceptor beads separately) Streptavidin Alpha Donor beads, unconjugated AlphaScreen Acceptor beads

unconjugated (no coating) Alpha Donor beads. Conjugate your own biomolecule to the AlphaScreen Acceptor beads to customize your own Alpha assay.

Streptavidin Alpha Donor beads for capture of any biotinylated molecule

Glutathione Alpha Donor beads for capture of any GST-tagged molecule

Nickel Chelate Alpha Donor beads for capture of any HIS6-tagged molecule

Instrumentation and Automation

AlphaScreen Conjugation Kit 2 x 2 mg 6760000K

Unconjugated Alpha Donor Beads 1 mg 6762013 5 mg 6762011 50 mg 6762012

Streptavidin Alpha Donor Beads 1 mg 6760002S 5 mg 6760002 50 mg 6760002B

Glutathione Alpha Donor Beads 1 mg 6765300 5 mg 6765301 25 mg 6765302

Nickel Chelate Alpha Donor Beads 1 mg AS101D 5 mg AS101M 25 mg AS101R

Contact your local PerkinElmer sales representative or visit www.perkinelmer.com/lifesciences

AlphaScreen Detection KitsProduct Information Product Description Size Part Number

AlphaScreen IgG (Protein A) AlphaScreen IgG (Protein A) Detection Kit 500 assay points 6760617C 10,000 assay points 6760617M 50,000 assay points 6760617R AlphaScreen Mouse IgG Detection Kit 500 assay points 6760606C 10,000 assay points 6760606M 50,000 assay points 6760606R AlphaScreen Rabbit IgG Detection Kit 500 assay points 6760607C 10,000 assay points 6760607M 50,000 assay points 6760607R

Anti-IgG (Protein A) AlphaScreen Acceptor beads, streptavidin-coated Alpha Donor beads, biotinylated-rabbit IgG, buffer (10X) Anti-mouse IgG (mIgG) AlphaScreen Acceptor beads, streptavidin-coated Alpha Donor beads, biotinylated-mouse-IgG, buffer (10X) Anti-rabbit IgG (rIgG) AlphaScreen Acceptor beads, streptavidin-coated Alpha Donor beads, biotinylated-rabbit-IgG, buffer (10X)

Product Information Product Description Part Number

Fusion Tag Anti-tag AlphaScreen Acceptor beads, streptavidin-coated Alpha Donor beads, biotinylated-tag, buffer (10X)

C = 500 assay points M = 10,000 assay points R = 50,000 assay points

1 tube of biotinylated-tag for use with AlphaScreen Fusion Tag Kits (GST and Histidine)

AlphaScreen GST (Glutathione-S-Transferase) 6760603C, 6760603M, 6760603R Detection Kit

AlphaScreen DIG (Digoxin/Digoxigenin) 6760604C, 6760604M, 6760604R Detection Kit

AlphaScreen FITC (Fluorescein) Detection Kit 6760605C, 6760605M, 6760605R

AlphaScreen c-Myc Detection Kit 6760611C, 6760611M, 6760611R

AlphaScreen HA (Hemagglutinin) Detection Kit 6760612C, 6760612M, 6760612R

AlphaScreen FLAG (M2) Detection Kit 6760613C, 6760613M, 6760613R

AlphaScreen Histidine (Nickel Chelate) Detection Kit 6760619C, 6760619M, 6760619R

Biotinylated-GST; 1.5 mL 6760305M

Biotinylated-HIS6; 500 µL 6760303M

www.perkinelmer.com/immunogenicity

ORDERING INFORMATION

Product Information Product Description Part Number

AlphaLISA Acceptor Beads(order Alpha Donor beads separately)

Protein A AlphaLISA Acceptor Beads AL101C, AL101M, AL101R

Protein G AlphaLISA Acceptor Beads AL102C, AL102M, AL102R

Anti-human IgG (Fc specific) AlphaLISA Acceptor Beads AL103C, AL103M, AL103R

Anti-rabbit IgG (Fc specific) AlphaLISA Acceptor Beads AL104C, AL104M, AL104R

Anti-mouse IgG (Fc specific) AlphaLISA Acceptor Beads AL105C, AL105M, AL105R

Anti-rat IgG (Fc specific) AlphaLISA Acceptor Beads AL106C, AL106M, AL106R

Anti-goat IgG (Fc specific) AlphaLISA Acceptor Beads AL107C, AL107M, AL107R

Nickel Chelate AlphaLISA Acceptor Beads AL108C, AL108M, AL108R

Glutathione AlphaLISA Acceptor Beads AL109C, AL109M, AL109R

Anti-GST AlphaLISA Acceptor Beads AL110C, AL110M, AL110R

Anti-c-Myc AlphaLISA Acceptor Beads AL111C, AL111M, AL111R

Anti-FLAG AlphaLISA Acceptor Beads AL112C, AL112M, AL112R

Anti-DIG AlphaLISA Acceptor Beads AL113C, AL113M, AL113R

AlphaLISA Acceptor Beads

C = 250 µg M = 5 mg R = 25 mg

Product Information Product Description Size Part Number

unconjugated (no coating) AlphaScreen Acceptor beads. Conjugate your own biomolecule to the AlphaScreen Acceptor beads to customize your own AlphaScreen assay.

Single Protein A-coated AlphaScreen Acceptor beads

Unconjugated AlphaScreen Acceptor Beads 1 mg 6762003 5 mg 6762001 50 mg 6762002

AlphaScreen Protein A Acceptor Beads 5 mg 6760137M 25 mg 6760137R

AlphaLISA Unconjugated Acceptor Beads 1 mg 6772001 5 mg 6772002 50 mg 6772003

unconjugated (no coating) AlphaLISA Acceptor beads. Conjugate your own biomolecule to the AlphaLISA Acceptor beads to customize your own AlphaLISA assay.

Alpha Toolbox (cont.)Product Information Product Description Size Part Number

AlphaScreen Acceptor Beads(order Alpha Donor beads separately)

DELFIA Plates and ReagentsProduct Information Package Product Number

ELAST ELISA Amplification KitsProduct Information Description Product Number

60 Plates AAAND-0001

10 Plates AAAND-0005

250 µg 1244-360

200 µg/1 mg AD0105/AD0106

50/250/1000 mL 1244-106/1244-111/4002-0010

250/1000 mL 1244-114/4010-0010

50/250/1000 mL 1244-104/1244-105/4001-0010

Contains reagents to amplify signals in ten 96-well ELISA microplates NEP116001EA

Contains reagents to amplify signals in one 96-well ELISA microplate NEP116E001EA

DELFIA Yellow Plate, 96-well

DELFIA Streptavidin-coated Yellow Plate, 96-well

DELFIA Eu-labeled Streptavidin

DELFIA Eu-labeled Goat Anti-rabbit IgG

DELFIA Assay Buffer

DELFIA Wash Concentrate

DELFIA Enhancement Solution

ELAST ELISA Amplification System

ELAST Evaluation Size

For a complete listing of our global offices, visit www.perkinelmer.com/ContactUs

Copyright ©2010, PerkinElmer, Inc. All rights reserved. PerkinElmer® is a registered trademark of PerkinElmer, Inc. All other trademarks are the property of their respective owners. 009247_01 Printed in USA Jul. 2010

PerkinElmer, Inc. 940 Winter Street Waltham, MA 02451 USA P: (800) 762-4000 or (+1) 203-925-4602www.perkinelmer.com

For scientific breakthroughs. For a better tomorrow.

At PerkinElmer, we share your commitment to answering challenging biological problems. Our broad portfolio of light-based detection solutions and expertise empower you to do more. We offer the right combination of technologies to deliver more physiologically relevant results and enhance your lab’s productivity and efficiency. With light-emitting chemistries, light-reading instruments and enlightening software, brilliant discoveries can shine through.

Contact us at 800 762-4000 (U.S.), 00 800 33 29 0000 (Europe) or 800 820 5046 (China). Or contact us by e-mail at global.techsupport@perkinelmer.com.

Learn more at www.perkinelmer.com/immunogenicity