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A Rapid, Flexible Alternative to ELISA for Protein

Detection, Characterization and Quantitation

Broadcast Date: Tuesday, September 25, 2012

Time: 11:00 am EDT

Sponsored by





Your Moderator

Tamlyn Oliver Managing Editor

Genetic Engineering & Biotechnology News



Francesco Lipari, Ph.D.

Senior Scientist

Biotherapeutic Applications

PerkinElmer Life Sciences and Technology

© 2009 PerkinElmer © 2009 PerkinElmer © 2009 PerkinElmer © 2009 PerkinElmer

Alpha Technology: Introduction Francesco Lipari, Ph.D. September 2012

Alpha Technologies (AlphaScreen, AlphaLISA)

Bead based proximity assay

Donor beads and Acceptor beads are

brought into proximity through

biomolecular interactions

The Donor beads are excited with a laser

at 680nm resulting in the release of singlet

oxygen

Singlet oxygen initiates an amplified

fluorescent signal cascade in the Acceptor

bead producing an emission of light

between 520 and 620nm

Emission occurs only if the beads are

brought within 200nm proximity

The magnitude of light emission is

proportional to the amount of biomolecules

interacting together

Si N

N

N

N

N

N

N

N

OSi

SiO

Alpha Technologies cover broad range of affinities

aM fM pM nM mM mM

Sugar-Lectin Protein-

Protein

Biotin-

Streptavidin

Receptor-

Ligand

Alpha

Filtration assays

FP

ELISA-like

TR-FRET

AlphaLISA protocol

A Rapid, Flexible Alternative to ELISA for Protein Detection, Characterization, and Quantitation

Michael Bembenek, Ph.D., Lead Discovery, Associate Scientific Fellow; Millennium:

The Takeda Oncology Company

“Determination of Complementary Antibody Pairs using Protein A Capture with the

AlphaScreen Assay Format”

Chamindie Punyadeera, Ph.D., Senior Research Fellow, Australian Institute for

Bioengineering and Nanotechnology, The University of Queensland

“Saliva Biomarker Detection Powered by AlphaLISA technology”

Francesco Lipari, Ph.D., Senior Scientist, Biotherapeutics Applications,

PerkinElmer Life Sciences and Technologies

“AlphaLISA: A Simple, Homogeneous Immunoassay Platform for Bioprocess Analytics”



Michael E. Bembenek, Ph.D. Lead Discovery

Scientific Fellow

Millennium: The Takeda Oncology Company

2012/9/17 Millennium: The Takeda Oncology Company

Michael E. Bembenek, Ph.D.

Determination of Complementary Antibody Pairs using Protein A Capture with the AlphaScreen Assay Format

Lead Discovery

Scientific Fellow

｜○○○○ | 09/2012 10


GOAL = HOW TO FIND ANTIBODY PAIRS QUICKLY FOR USE IN BIOMARKER DISCOVERY

Benefits of complementary pairing antibodies include:

– use of sandwich format improves selectivity and specificity for biomarker.

– ability to move away from Western blot based assay methods because of limitations with quantitation & labor intensity.

No current high throughput method for identifying complementary antibody pairs.

｜○○○○ | 09/2012 11


DETERMINATION OF COMPLEMENTARY ANTIBODY PAIRS USING PROTEIN A CAPTURE WITH THE ALPHASCREEN ASSAY FORMAT*

Describe a new method for identifying complementary antibody

pairs derived from commercial sources.

Describe method for identifying primary antibodies from rabbit

monoclonal hybridoma selection using “tags” on recombinant

target proteins and then,

Identify complementary antibody pairs during the hybridoma

screening process – illustrate with 2 examples.

Use of complementary antibody pairs for quantitation of biomarker.

｜○○○○ | 09/2012 12

* Michael E. Bembenek, Anne Burkhardt, Jingya Ma, Zhi Li, Huay-keng Loke, Dongyun Wu,

Qing Xu, Olga Tayber, Liying Xie, Ping Li, Li Li. Determination of complementary antibody pairs

using protein A capture with the AlphaScreen assay format.

Analytical Biochemistry, 2011, 408(2), 321-327.


BASICS OF EXPERIMENTAL DESIGN

Need to prepare the Protein A donor and acceptor

AlphaScreen beads harboring captured antibodies

then

Design a matrix to survey the antibody pairings.

｜○○○○ | 09/2012 13

Emission

520-620 nm

AlphaScreen

Acceptor

Bead

1O2

Excitation

680 nm

AlphaScreen

Donor Bead

200nm

Principle of AlphaScreen Technology


Finding the Complementary Pairs

｜○○○○ | 09/2012 14

Uses Protein A on BOTH donor & acceptor beads to find

complementary pairs Ξ postive AlphaScreen signal

In order to

achieve this a

matrix

experiment is

designed ……


Proof of concept example = screen 3 commercial pan anti-Akt

Antibodies from Cell Signaling, Inc.

｜○○○○ | 09/2012 15

Protein A Donor bead

Protein A Acceptor bead

Coat each bead type with individual antibody separately

WASH Beads with buffer to remove excess unbound Ab’s

Collect Each Individual Antibody Coated Bead Type By Centrifugation

Resuspend each Ab coated bead then cross-titrate on a matrix in presence of recombinant Akt2


“AlphaMatrix” for Finding Complementary Pairs

｜○○○○ | 09/2012 16

3 X 3 Matrix

pan-Akt antibody

(CST:#4685)

on

Protein A Donor

pan-Akt antibody

(CST:#4691)

on

Protein A donor

pan-Akt

polyclonal

antibody

(CST:#9272)

on

Protein A donor

pan-Akt antibody

(CST:#4685)

on

Protein A

acceptor

Same Ab

against itself

pan-Akt antibody

(CST:#4691)

on

Protein A

acceptor

Same Ab

against itself

pan-Akt polyclonal

antibody

(CST:#9272)

on

Protein A acceptor

Same Ab

against itself

Pairs are represented TWICE

on the matrix on different bead

types = off the diagonal

Each column

contains an equal

aliquot of

antibody coated

onto Protein A

donor beads

Each row contains an

equal aliquot of antibody

coated onto Protein A

acceptor beads


“AlphaMatrix” for Finding Complementary

Total Akt Pairs: the Data

｜○○○○ | 09/2012 17

17

pan-Akt antibody

(CST:#4685)

on

Protein A Donor

pan-Akt antibody

(CST:#4691)

on

Protein A donor

pan-Akt polyclonal

antibody

(CST:#9272)

on

Protein A donor

pan-Akt antibody

(CST:#4685)

on

Protein A acceptor

3439 3363 86906

pan-Akt antibody

(CST:#4691)

on

Protein A acceptor

5890 15124 459515

pan-Akt polyclonal

antibody

(CST:#9272)

on

Protein A acceptor

54815 306242 15409

3 X 3 Matrix

Millennium: The Takeda Oncology Company ｜○○○○ | 09/2012 18

Useful for Quantitating Total Akt2 in a

Antibody Sandwich Format

pan-Akt antibody

(CST:#4691)

on

Protein A acceptor beads

vs

pan-Akt polyclonal antibody

(CST:#9272)

on

Protein A donor beads


HIGHER THROUGHPUT HYBRIDOMA ANTIBODY SCREENING

Use method for identifying positive rabbit hybridoma producing

antibodies to recombinant “tagged” protein antigens.

Select positive hybridoma clones with verification by Western blot &

immunoprecipitation.

Capture positive antibodies from hybridoma derived cell culture

supernatants onto both Protein A donor and acceptor beads –

requires no tagging of Ab’s like many other methods.

Use “AlphaMatrix” format to identify complementary pairs and

for final monoclonal selection.

｜○○○○ | 09/2012 19


The Antibody Hunt Example #1: identifying positive rabbit hybridoma antibodies with

AlphaScreen for ATF3

｜○○○○ | 09/2012 20

Identify positive hybridoma clonal supernatants by using a tagged recombinant antigen

as bait, i.e., GST-ATF3, in AlphaScreen format.

Glutathione

Donor Bead Antibody Captured

From Hybridoma

Supernatant

Protein A Acceptor Bead

Choose top seven positive clones

based upon AlphaScreen

detection Method.

Assay time is less than 3 hrs.

GST-ATF3


Confirmed by Western Blot & Immunoprecipitation

｜○○○○ | 09/2012 21

Western

blot of

transiently

transfected

293T cells

vs

hybridoma

clones(#’d).

Immunoppt. of

transiently

transfected

293T cell lysates vs

hybridoma

clones(#’d).


Results of “AlphaMatrix” for Finding Complementary

Pairs to ATF3 for the 7 positive hybridomas

｜○○○○ | 09/2012 22

Using an arbitrary cut-off of ~6000 CPS for the average of the positive pairings

listed in the 7 X 7 matrix highlighted in yellow boxes.

The antibodies from hybridoma clone 38 showed the highest level of pairings with

five of six potential complementary monoclonal antibodies followed by clone 19

which paired with three of six possible clones.

The three highest ranking pairs were selected for subcloning and expansion.


Example #2: Identifying positive hybridomas with AlphaScreen

for the b subunit of Neddylation activating enzyme(NAE)

｜○○○○ | 09/2012 23

Top 20 hybridoma clones based

upon AlphaScreen detection

method are listed in yellow

highlighted boxes.

Western blot & IP positives

reduced to the number to18.

Antibody captured from

hybridoma supernatant

Anti-Flag

Acceptor bead

Protein A

donor

bead

FLAG-NAEβ

Positives were then screened in an 18X18 AlphaMatrix for identifying complementary pairs


Results of “AlphaMatrix” for Finding Complementary

Pairs to NAEb via 18 X 18 Matrix

｜○○○○ | 09/2012 24

Clones 18, 81 & 87 produced best pairing antibodies


Picking the Best Signal Producing Subclones for the

Anti-NAEb Pairs

｜○○○○ | 09/2012 25

All subclones produced positive complementary antibodies from initial

screen as illustrated with example shown below with clones 81 vs 87.

Rescreen by “AlphaMatrix” identified top hybridoma clones with best

overall signal for the pairs of hybridoma subclones 81 and 87.

Top three subclones were used for scale-up and antibody production as

highlighted in yellow boxes.


Quantitative - Using the Antibody Pairs for

Standard Titration Curves

｜○○○○ | 09/2012 26

Complementary antibodies captured onto Protein A donor & acceptor beads.

Both curves show mass capture limits with a “hook” above 1 nM and near linear

responses below 0.5 nM analyte with a lower limit of detection in the pM range.


Method Allows Detection and Selection of Complementary

Antibody Pairs

Assay is rapid & high throughput – requires no antibody tagging.

Can determine complementary antibody pairs from commercial

source antibodies, i.e, total Akt.

Also useful for hybridoma screening and antibody development,

for example, ATF3 & NAEb.

Translates to useful and sensitive sandwich based AlphaScreen

format for direct quantitation using both in vitro cell-based

assays and pharmacodynamic biomarkers in xenografts.

｜○○○○ | 09/2012 27


Collaborators Reference:

Michael E. Bembenek, Anne Burkhardt,

Jingya Ma, Zhi Li, Huay-keng Loke,

Dongyun Wu, Qing Xu, Olga Tayber,

Liying Xie, Ping Li, Li Li. Determination

of complementary antibody pairs using

protein A capture with the AlphaScreen

assay format. Analytical Biochemistry,

2011, 408(2), 321-327.

MPI - Anne Burkhardt, Jingya

Ma, Zhi Li, Huay-keng Loke,

Dongyun Wu, Qing Xu, Olga

Tayber, Ping Li

Epitomics, Inc. - Liying Xie and

Li Li

Special thanks to Mike Kuranda &

William Mallender

for helpful scientific discussions.

｜○○○○ | 09/2012 28

2012/9/17

We aspire to cure cancer



Chamindie Punyadeera, Ph.D. Senior Research Fellow

The University of Queensland Diamantina Institute

The Translational Research Institute Pty Ltd.

Chamindie Punyadeera (PhD)

Email: [email protected]

Saliva Biomarker Detection Powered

by AlphaLISA® Technology

Outline Saliva vs. blood

Human saliva

Saliva production and functions

Molecular transportation from blood into saliva

Salivary proteome

Biomarker applications using AlphaLISA®

Technology

Early detection of heart disease (Ischemic Heart Disease)

Early detection of head and neck cancers

Saliva vs Blood Blood Saliva

Invasive. Non-invasive, cost effective.

Requires processing to

separate cells from serum.

No processing is needed.

Requires skilled people to

collect blood i.e. doctors and

nurses.

Anyone can collect a saliva

sample, enabling multiple

collections within a day.

A high risk of contracting

infectious pathogens.

Minimal/low risk of

contracting pathogens.

Not so ideal due to costs. Ideal for 3rd world and remote

communities.

Analytes present at medium to

high levels.

Analytes present at low levels

(pg/mL).

Functions of saliva (adapted from M.J. Levine, 1993)

Functions

Anti-

Bacterial Buffering

Digestion

Mineral-

ization

Lubricat-

ion Tissue

Coating

Anti-

Fungal

Anti-

Viral

Carbonic anhydrases,

Histatins

Amylases,

Mucins, Lipase

Cystatins,

Histatins, Proline-

rich proteins,

Statherins

Mucins, Statherins

Amylases, Statherins

Cystatins, Mucins,

Proline-rich proteins,

Histatins

Cystatins,

Mucins

Amylases, Cystatins,

Histatins, Mucins,

Peroxidases

adapted from M.J. Levine, 1993

Molecular transportation from blood into saliva

Pfaffe et al.,

Clin.Chem,

March 2011

Salivary proteome (# 2340 proteins)

(E Scarano

et. al., Acta

Otorhinolaryn

gol Ital. 2010)

AlphaLISA® is an Easy to use, No- Wash and

highly reproducible Platform

Current protocol AlphaLISA ®

How do we use AlphaLISA® to

detect salivary proteins

1) Determine the salivary matrix effects on the

assay

2) Develop assays using 10mL assay reaction

vs. 50mL and determine assay intra-assay CV

3) Determine the assay efficiency for shorter

incubation times 90 minutes vs. 15 minutes

Matrix effects: Salivary C-Reactive

protein assay

Salivary C-Reactive Protein assay reproducibility

Incubation 90 mins. vs 15 mins.

Salivary CRP to detect Ischemic Heart Disease

Salivary CRP levels vs blood levels

Pearson correlation = 0.9

A

Drool method (passive)

Saliva collection methods (Topkas et al., Clinica Chimica Acta, March, 2012)

Salimetrics® Oral Swab (stimulated)

Salivette® (Stimulated)

Greiner Bio-One (Stimulated)

Dro

ol

SOS

Sal

ivet

te® C

otton

Sal

ivet

te® S

ynth

etic

0

50

100

150

200

250

*

C-R

eacti

ve p

rote

in

(pg

/ml)

Dro

ol

SOS

Sal

ivet

te® C

otton

Sal

ivet

te® S

ynth

etic

0

200

400

600

800

*

Imm

un

og

lob

in E

(p

g/m

l)

Dro

ol

SOS

Sal

ivet

te® C

otton

Sal

ivet

te® S

ynth

etic

0

200

400

600

800

** *

Myo

glo

bin

(p

g/m

l)

Figure 3

Salivary myoglobin, CRP and IgE levels (Topkas et al., Clinica Chimica Acta, March, 2012)

The use of AlphaLISA® Epigenetic assays in Head and Neck Cancer

Hard facts: head and neck cancers

Approximately 780,000 (p/a) new cases of head and neck

squamous cell carcinomas (HNSCC) diagnosed world wide and

300,000 deaths each year. (Boyle P. et al. World Cancer Report 2008).

Tobacco use is a major risk factor for HNSCC (other factors

diet, mouth wash use and HPV infections) (Spitz and Trizna, 1999).

Smoking kills over 1,000,000 people (p/a), causing 30% of all

cancer related deaths. Yet, 1 in 3 people, world-wide is addicted

to nicotine.

The direct impact of smoking can clearly be seen in the oral

cavity due to its proximity, thus, human saliva is ideal as a

detection medium.

Epigenetics and Cancer • Epigenetic regulation is critical

for mammalian development and

cellular differentiation.

• DNA methylation/Histone

modification in cells is one of the

earliest events to occur during

cancer initiation and has

demonstrated utility:

Early disease detection

Better disease stratification

Predicting disease relapse

Response to therapy (Esteller, M. Epigenetics in Cancer, New England Journal of

Medicine, 358: 2008).

Future work using AlphaLISA

assays for cancer epigentics studies

The AlphaLISA Epigenetic Cellular Detection Kits in

cancer enable rapid and direct detection of endogenous

modification of epigenetic markers on histones H3 and

p53.

AlphaLISA cellular epigenetic assay advantages over

westerns/ELISA:

• Simple assay protocol which can be automated

• Suitable for endogenous and recombinant cell lines,

providing the flexibility to work with relevant cell models

Summary

• Advantages of AlphaLISA® technology over standard

immunoassays:

- AlphaLISA® is applicable with complex biological

fluids such as saliva with careful sample preparations

- Amenable to point of care assay platforms.

- Easy to use and optimize the assay to eventually

meet diagnostic assay quality for clinical research.

- Cost effective

• One of the disadvantages of AlphaLISA® is the

inability to multiplex.

Acknowledgments Saliva Team (present)

• Michael Caragata

• Yunxia Wan

• Prati Pandit

• Jared Foo

• Rahul Nagadia

• Alicia

• Pengxiang Ji

• Bob Xi Zhang

Past: •Tina Pfaffe

• James Dkhar

• Thea Cullen

• Eleni Topkas

• Patricia Keith

• Rosalinda Mohamed

• Jennifer

•Ling Li Long

SPIT Collaborators:

• Professor Justin Cooper white (UQ)

• Professor William Coman (UQ)

• Head and Neck Cancer clinic staff

• Dr Paul Slowey (Oasis Diagnostics, USA)

• Dr John Duley (UQ)

• Professor Ian Frazer (UQDI/TRI)

• Professor Matthew Cooper (UQ)

• Professor Karam Kostner (UQ/Mater)

• Professor John Atherton (RBWH/UQ)

• Dr Ben Schultz (UQ)

• Dr Goce Dimeski (PAH)

• Dr Dmitry Ovchinnikov (UQ)

• Professor David Wong (UCLA/USA) Research funding:

• Queensland Government Strategic Funds

• The University of Queensland Research Excellence Award

• University of Queensland Collaborative Industry Engagement

“The only true wisdom is not knowing nothing”

Socrates




Senior Scientist



54 54 © 2009 PerkinElmer © 2009 PerkinElmer © 2009 PerkinElmer © 2009 PerkinElmer

AlphaLISA: A Simple, Homogeneous Immunoassay Platform for Bioprocess Analytics Francesco Lipari, Ph.D. September 2012

55 55

Advantages of AlphaLISA technology

Assays for the bioprocess lab

Outline

56 56

Trends in bioprocess analytics

More high throughput requirements due to the implementation of high throughput cell

culture and purification technologies

Homogeneous AlphaLISA assay allows:

Consistent, reproducible results - %CVs less than 10% (intra-assay)

Very simple protocol and easy automation- just 4 steps to results

Low affinity interactions are not washed away

Complete assay in < 3 hours

Sensitivity is similar to classical ELISA technology, but without wash steps

Low sample volumes (≤ 10 mL)

AlphaLISA Technology Advantages

57 57

The AlphaLISA Assays for Bioprocess Development

Quantitation of monoclonal antibody

Human IgG

Detection of process contaminants

Human albumin

Residual Protein A

CHO Host Cell Proteins

PER.C6 Host Cell Proteins

NS0 Host Cell Proteins

E. Coli Host Cell Proteins

58 58

High throughput cell culture and protein purification development in 96-well

format generates hundreds of samples for determination of monoclonal

antibody titer.

Alpha technology provides an ideal high throughput technology to analyze

many samples in a short period of time.

Large dynamic range (0.2 – 300 ng/mL)

Human IgG Assay

1,000

10,000

100,000

1,000,000Buffer

DMEM

DMEM/F12

MEM

RPMI

-12 -11 -10 -9 -8 -7 -6-

Log [hIgG] (g/mL)

Alp

ha

LIS

A S

ign

al (c

ou

nts

)

59 59

Cell culture medium used to produce biotherapeutics may contain human albumin.

A highly sensitive assay is required to determine the residual levels of albumin

remaining during the processing steps.

Highly sensitive: LDL = 110 pg/mL

Company 1 ELISA kit: LDL = 150 pg/mL

Company 2 ELISA kit: LDL = 250 pg/mL

Precision

Intra-assay

Inter-assay

Human Albumin Assay

Sample Mean (pg/mL) SD (pg/mL) %CV (n=18)

A 34310 2144 6.2

B 10358 533 5.1

C 1117 123 11.0

Sample Mean (pg/mL) SD (pg/mL) %CV (n=6)

A 34310 4498 13.1

B 10358 1068 10.3

C 1117 151 13.5

60 60

Residual Protein A assay

Protein A is leached from protein A columns used for the initial purification step in

monoclonal antibody production. It is particularly challenging to quantify protein A

in samples due to the presence of antibody at a high concentration.

Standards or samples containing less than or equal to 1mg/mL of IgG

Add 10mL of 3X dissociation buffer to 20mL of standard or sample

Incubate 60 min at 98 oC (PCR instrument)

Centrifuge for 5 mins at greater than or equal to 200g

Perform standard AlphaLISA protocol

61 61


-9 -8 -7 -6 -5 -4 -3 -20

25,000

50,000

75,000

100,000RT

Heat Treated

Log [hIgG] (g/mL)

Alp

ha

LIS

A S

ign

al (c

ou

nts

)

Importance of heat treatment

Sensitivity and selectivity

LDL = 11 pg /mL

Similar cross-reactivity to Protein A used in MabSelect SuReTM, MabSelectTM &

MabSelect XtraTM

62 62


Buffer Spike

(ng/mL) % Recovery

PBS 0.5X, 0.5 M NaCl

3.0 88

0.3 86

0.03 79

Tris 50 mM, pH 8.0

3.0 91

0.3 92

0.03 96

Citrate / Phosphate pH 6.0

3.0 116

0.3 94

0.03 102

Phosphate buffer 50 mM, pH 6.0

3.0 92

0.3 92

0.03 104

Acetate buffer, pH 5.0

3.0 110

0.3 99

0.03 102

Suitable for different buffers

63 63

Removal of host cell proteins (HCPs) during bioprocessing is key to providing a

safe biotherapeutic, due to the potential immunogenicity of the foreign proteins.

Assays for measuring HCP must be highly sensitive and applicable to a variety of

purification buffers.

Difference between original CHO HCP kit and new kit – different antibodies that

should detect more CHO proteins. Ideally should compare the two kits and choose

most appropriate for the process.

Host Cell Protein Assays

HCP assay Lower Detection Limit

(ng/mL)

Upper Limit

(mg/mL)

E. coli HCP 0.5 1

NS0 HCP 1.6 1

CHO HCP 0.2 0.3

CHO HCP (broad

reactivity) 0.5 1

PER.C6 HCP 0.8 1

64 64

CHO HCP (broad reactivity) assay - test different cell culture media


100

1,000

10,000

100,000

1,000,000

-11 -10 -9 -8 -7 -6 -5

HiBlock Buffer

-

RPMI + 10% FBS

Hybridoma-SFM

CD Hybridoma Medium

Log [Analyte] (g/mL)

Alp

ha

LIS

A S

ign

al (c

ou

nts

)

65 65


0.0

0.5

1.0

1.5

2.0

2.5

3.0

0

20

40

60

80

100

0 50 100 150 200 250 300

Alpha HCP Kit: 2012.06.25

[Ho

st C

ell

Pro

tein

] (

mg/

mL)

Ce

ll Viab

ility (%)

Cultivation Time (h)

0.0

0.5

1.0

1.5

2.0

2.5

3.0

0

20

40

60

80

100

120

140

0 50 100 150 200 250 300

Alpha HCP Kit: 2012.06.25

[Ho

st C

ell

Pro

tein

] (

mg/

mL)

Viab

le ce

lls (x10

6 cells/mL)

Cultivation Time (h)

Professor Omasa’s laboratory, The Univeristy of Tokushima, Institute of Technology

and Science

66 66


E. coli HCP assay - suitable for different buffers

Buffer Spike

(ng/mL)

%

Recovery

%Recovery (1/2

diluted samples)

PBS with 1 mg/mL mouse IgG, pH

7.2

100

10

54

50

72

68

PBS with 10 mg/mL BSA, pH 7.2 100

10

72

75

94

83

0.037 M Citrate/0.13 M Phosphate

with 1 mg/mL BSA, pH 6.0

100

10

61

50

78

63

0.1M Acetate, 10mg/mL BSA, 1%

Triton, pH 5.0

100

10

62

44

104

70

0.05 M Tris, 10mg/mL BSA, pH 8.5 100

10

78

84

119

99

67 67

Alpha technology offers:

Consistent, reproducible results - %CVs less than 10%

Simple protocol - just 4 steps to results in < 3 hrs

Equal or better sensitivity than other immunoassay technologies without tedious wash

steps

Suitable for a variety of bioprocess sample matrices

Wide dynamic range - up to 4 logs

Use anywhere you are currently using ELISA or electrochemiluminescent (ECL)

techniques

Conclusions



A Rapid, Flexible Alternative to ELISA for Protein Detection,

Characterization and Quantitation

Q&A



Your Moderator

Tamlyn Oliver Managing Editor

Genetic Engineering & Biotechnology News




Senior Scientist





Michael E. Bembenek, Ph.D. Lead Discovery

Scientific Fellow




Chamindie Punyadeera, Ph.D. Senior Research Fellow

The University of Queensland Diamantina Institute

The Translational Research Institute Pty Ltd.



Thank You For Attending

A Rapid, Flexible Alternative to ELISA for Protein Detection,

Characterization and Quantitation

Broadcast Date: Tuesday, September 25, 2012

Time: 11:00 am EDT

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