adcc reporter bioassay

Promega Corporation Promega Corporation ©2013 Promega Corporation.

March 2014

ADCC Reporter Bioassays - V and F Variants: Novel, Bioluminescent Cell-Based Assays for Quantifying

Fc Effector Function of Antibodies

Rev 01

©2013 Promega Corporation.

Promega Corporation Promega Corporation 2

Topics Presented

Introduction to ADCC

– Problem with classic ADCC assays

Solution: Designing a Better ADCC Bioassay

– Introducing ADCC Reporter Bioassay

“Cells as Critical Reagents”

– Cell selection, frozen, thaw-and-use format

V Variant ADCC assay performance

– Assay optimization, qualification, potency & stability

indicating, benchmarking

F Variant ADCC assay performance

– Assay optimization, qualification, potency & stability Indicating

Commercial Formats

– Kits & Cell Propagation Model, custom materials

Image source:

www.iconplc.com

http://www.iconplc.com/



Reflective of the mechanism of action (MOA) of the

biological product

Well controlled (precise, accurate, robust, reproducible)

Stability-indicating

Usable as a QC lot-release assay

Modified from Chana Fuchs (DMA/CDER)

An Ideal Bioassay Is…

On the following pages, we demonstrate how the novel ADCC Reporter Bioassay fulfills each of these parameters



Characteristics of a Valid Bioassay Are…

-10 -9 -8 -7 -6 -50

5

10

15

20

25

30

35plate1

plate2

plate3

plate4

Log10 [B1 antibody], g/ml

Fo

ld o

f In

du

cti

on

Repeatability

Design: Two analysts Three days Four plates per day 100% vs 50% 100% vs 75% 100% vs 125% 100% vs 150% -10 -9 -8 -7 -6 -5

0

5

10

15

20

25

30

35

100%

50%

150%


Fo

ld o

f In

ductio

n

Y=1.026X-5.126 R2=0.995

Relative Potency

Linearity

Log [control antibody], g/ml

Log [control antibody], g/ml

Validation of Analytical

Procedures:

Accuracy

Precision: Repeatability (intra-assay

precision)

Intermediate precision (day

to day, analyst to analyst)

Reproducibility (lab to lab)

Specificity

Linearity

Range

Robustness

- ICH Guideline Q2 [R1]


March 2014

The Problem with Classic ADCC Assays



Classic ADCC Assays Are Limited by

The performance of…

Effector cells:

PBMCs (peripheral blood

mononuclear cells)

NK from PBMCs

NK cell lines

Target cells:

Load with chromium-51 or Eu

Monitor cell lysis (LDH, Calcein AM,

GAPDH, CytoTox-Glo™ Assay)

CytoTox-Glo™ Cytotoxicity Assay

Target

cell Y Primary

NK effector

cell Antibody

FcgRIIIa

Classic assay principle

Cell lysis


March 2014

Solution: A Better ADCC Bioassay

Introducing ADCC Reporter Bioassay



Image source: Leibson-PJ, Immunity (1997) 6(6): 655-661; doi: 10.1016/S1074-7613(00)80441-0

Luciferase reporter is

readout of pathway

activation state

Target-cell bound Ab binds to FcgRIIIa

(CD16) on effector cell – activating

pathway

Scientific Basis of ADCC Reporter Bioassay

New reporter-based bioassay measures a

step earlier in the pathway



Target

cell Y

= NFAT-RE-luc

FcgRIIIa

(V158 or

F158)

Antibody

Improving Upon the Classic ADCC Bioassay

Effector cell

(engineered

Jurkat)

Reporter-based ADCC bioassay

Glo

Target

cell Y Primary

NK effector

cell Antibody

FcgRIIIa

Classic ADCC bioassay

Specific signal is from target cell:

High variability of assay results –

mainly due to primary NK cells

Spontaneous lysis of target & effector

cells results in high background

Complex & tedious to perform

Signal is from effector cell:

Reduced variability - replace NK cells

with genetically engineered stable cell line

ADCC MOA-based bioassay

Simple & easy to perform

Improved bioassay performance with

robust reagents and assay design

Cell lysis

Parekh, BS et al (2012). mAbs, 4(3), 310-318; doi: 10.4161/mabs.19873



Why an F Variant Version of the Assay is Needed

Polymorphism in the FcgRIIIa Receptor Impacts ADCC Response

FcyRIIIa

158F/V or F/F 158 V/V

>85% population ~10-15% population

Less efficient Ab

binding and ADCC

More efficient Ab

binding and ADCC

The FcgRIIIa receptor has a

polymorphism at amino acid 158 with

human genotypes of VV, FV and FF. The

V variant receptor has higher affinity for

Abs. Patients with FV or FF genotype

respond less well to Ab drugs having

ADCC MOA.

Many new generation mAb drugs are being designed to improve Ab

affinity for the F variant of the receptor, to improve patient response.

Drug developers need a low variability ADCC bioassay specifically to

measure the new drug potency via the F158 variant of FcgRIIIa, and

regulatory agencies are asking for this information.

See the section beginning on page 34, “Polymorphism in the FcgRIIIa Receptor” for information on our newly developed

F Variant ADCC Reporter Bioassay.

V158

or

F158


March 2014

“Cells As Critical Reagents”



Cells as Critical Reagents

Traditional cell-based assay using fresh cells from cell culture (1-2 weeks)

Cell culture Cell count, harvest, prepare for assay

Thaw for culture

Read plates

New assay format using frozen, thaw-and-use cells: convenience and improved

run-to-run reproducibility

Thaw-and-Use Resuspend in assay buffer, plate for assay

Read plates

Time: 1-2 weeks

Time: <24hr See pages 50-51 for

information on a convenient, low cost luminometer from

Promega: GloMax® Discover



No Cell Culture Required

ADCC Reporter Bioassay features Frozen, Thaw-and-Use Cells

1. Human cell lines (Jurkat, WIL2-S, Raji) - Developed for immediate thaw-and-use in bioassay

- Designed for good recovery and robust response upon thawing

2. Thaw-and-Use format - Cell propagation conditions & defined freezing protocol control assay

performance for a consistent bioassay response

- No pre-culturing prior to assay means less variability introduced

- Indefinite storage

- Identical cells in bioassay, day to day

3. Minimizes pre-assay planning, time & labor - Ample cell banks provide long-term supply

means no

cell culture required

Thaw-and-Use cells

+



Biological Performance Equivalent to Fresh Cells

Fresh cells from continuous culture Frozen, thaw-and-use cells

EC50=3.5ng/ml FI=34-fold

EC50=2.9ng/ml FI=41-fold

Assay specifics:

E:T ratio = 6:1; fresh WIL2-S cells as target cells;

Bio-Glo™ reagent; 20hr induction for fresh Jurkat cells

assay; 6hr induction for frozen Jurkat cells assay.



Frozen, Thaw-and-Use WIL2-S Target Cells Provide Convenience

Kit Control Ab Rituximab

ADCC Reporter Bioassay response to ADCC Bioassay Control Antibody (left) or Rituximab

(right). The EC50 response using frozen, thaw-and-use WIL2-S cells was 16.8ng/ml for Control Ab,

Anti-CD20. For Rituximab, 1.94ng/ml.

Assay specifics:

E:T ratio = 6:1; 6hr induction; Bio-Glo™ reagent



Frozen, Thaw-and-Use Raji Target Cells, Too…

ADCC Reporter Bioassay response to ADCC Bioassay Control Antibody (left) or Rituximab

(right). The EC50 response using frozen, thaw-and-use Raji cells was 59.7ng/ml for Control Ab,

Anti-CD20. For Rituximab, 17.0ng/ml.

Kit Control Ab Rituximab

Assay specifics:

E:T ratio = 6:1; 6hr induction; Bio-Glo™ Reagent



Complete QC on Cells Ensures Consistent Results

Production cell batches are rigorously tested:

STR analysis – cell ID profile (human)

CO1 analysis (cytochrome oxidase) – test

for presence of species (human and other

potential contaminants)

Cell doubling time under propagation

conditions

Mycoplasma (Hoechst and direct culture)

Sterility

Cell density

Cell viability after thaw

Fill volume

ADCC Reporter Bioassay (EC50 and fold induction)


March 2014

Performance of ADCC Reporter Bioassay (V Variant)



Assessing Critical Assay Parameters

Induction time E:T ratio (constant Effector cell #)

Other parameters tested:

Assay buffer: serum concentration, use of low IgG serum

Cell numbers per well

Pre-plating and incubation time: target cell plating, antibody/target cells incubation

White flat-, V- or U-bottom assay plates



Bioassay Development Using DOE

Target cell / Ab Jurkat cell Target cell

run induction time hr incubation time(mins) plating number (K) plating number (K)

1 5.5 30 75 10

2 5.5 30 75 12.5

3 5.5 30 90 12

4 5.5 30 90 15

5 5.5 45 75 10

6 5.5 45 75 12.5

7 5.5 45 90 12

8 5.5 45 90 15

9 6 30 75 10

10 6 30 75 12.5

11 6 30 90 12

12 6 30 90 15

13 6 45 75 10

14 6 45 75 12.5

15 6 45 90 12

16 6 45 90 15

Variables:

Induction time

Target/Ab pre-incubation

Effector cell number

Target cell number

Outputs & Results: Good response (fold induction) = 19-27

Good (low) L-term* values = 0.1-0.2

*L-term is a measure of assay precision around the EC50

determination (log width of the 95% confidence interval around

logEC50)



Parallelism and measurement of Potency relative to the reference antibody

Linearity & Accuracy of observed versus expected potencies across the

desired working range of potencies

Precision

- intra-assay

- intermediate (inter-assay) precision

Specificity to show response is dependent on specific antibody and the

presence of target cells and FcgRIIIa on effector cells, and not other

components

Stability-indicating to show the bioassay is capable of detecting loss of

structural integrity of an antibody

These qualification studies are critical to demonstrate a useful and effective ADCC bioassay

Useful and Effective ADCC Bioassay Demonstrated

Qualification Studies:



V Variant: Able to Measure Potencies of On-Market mAb Biologic Drugs

Rituximab, anti-CD20, chimeric

IgG1. Approved to treat B-cell

non-Hodgkin lymphoma and

chronic lymphocytic leukemia.

Trastuzumab, anti-HER2,

humanized IgG1. approved to

treat HER2-positive breast

cancer and stomach cancer.

Cetuximab, anti-EGFR,

chimeric IgG1. Approved to treat

colorectal cancer and certain

types of head and neck cancer.

Panitumumab, anti-EGFR,

human IgG2 with NO ADCC.

Three best-selling, on-market mAb biologic drugs that posses ADCC as main MOA

Rituximab tests performed using ADCC Reporter Bioassay, Complete Kit; Trastuzumab and Cetuximab using Core kit. Both V Variant.



Able to Measure Fc Effector Function in ADCC for TNF Blockers

mTNF CHO-K1 Target cells

Y = NFAT-RE-luc

FcgRIIIa

anti-TNF

engineered Jurkat

effector cells

Glo

mTNF

Assay specifics:

Effector cells: ADCC Bioassay Effector Cells,

(V Variant) frozen, thaw-and-use

Target cells: mTNF CHO-K1 target cells

E:T ratio: 7.5:1

An engineered CHO-K1 cell line

expressing membrane-bound TNF was established as target cells.

Infliximab

Adalimumab



Understanding Potency Determinations Using Quantitative Bioassays

Relative Potency: Use a bioassay to establish potency activity of unknown

biologic relative to a reference standard

Relative potency can only be determined when: The upper and lower asymptotes as

well as the slopes of the curves are not significantly different. Hence the curves are parallel

Only the EC50s differ

Relative potency calculation:

__EC50 Test Sample___ EC50 Reference Sample

y = d +a - d

1 + (conc/c)b

a

c

b

d

Concentration

Response

Potency (% of Reference)

4-parameter logistic curve fit and potency determination

Reference

Test sample

upper asymptote

slope

lower asymptote

EC50



Assay Qualification Results with WIL2-S Cells

Design: •Two analysts• Three days• Four plates per day100% vs 50%100% vs 75%100% vs 125%100% vs 150%

Linearity

Representative plate layout

Repeatability

Y=1.026X-5.126R2=0.995

1 2 3 4 5 6 7 8 9 10 11 12 Plate1

A

B no Ab dilu9 dilu8 dilu7 dilu6 dilu5 dilu4 dilu3 dilu2 dilu1 100%

C no Ab dilu9 dilu8 dilu7 dilu6 dilu5 dilu4 dilu3 dilu2 dilu1 50%

D no Ab dilu9 dilu8 dilu7 dilu6 dilu5 dilu4 dilu3 dilu2 dilu1 100%

E no Ab dilu9 dilu8 dilu7 dilu6 dilu5 dilu4 dilu3 dilu2 dilu1 50%

F no Ab dilu9 dilu8 dilu7 dilu6 dilu5 dilu4 dilu3 dilu2 dilu1 100%

G no Ab dilu9 dilu8 dilu7 dilu6 dilu5 dilu4 dilu3 dilu2 dilu1 50%

H

Precision = average of RSD (%) = 7.3%

Accuracy= average of Recovery (%) = 95.8%

Antibody

Sample

Measured

Potency

(%)

Mean

Potency

(%) SD %

Recovery

(%)

RSD

(%)day 1 48.5

day 2 50% 45.2 48.9 3.9 97.7 7.9

day 3 52.9

day 1 63.1

day 2 75% 62.9 66.4 5.9 88.5 8.9

day 3 73.2

day 1 112.1

day 2 125% 136.3 123.0 12.3 98.4 10.0

day 3 120.5

day 1 148.8

day 2 150% 150.4 147.6 3.6 98.4 2.4

day 3 143.6

Bioassay uses frozen, thaw-and-use cells for both effector & WIL2-S target cells

Good repeatability, accuracy, precision and linearity were obtained



Assay Qualification Results with Raji Cells

DayAntibody Sample

Measured Potency

(%)

Mean Potency

(%)

SD (%)

Recovery (%)

CV (%)

1 49.9

2 50% 51.3 51 0.7 102 1.4

3 50.5

1 78.9

2 75% 78.8 76 5.11 101 6.73 701 118.6

2 125% 116.9 117 1.19 94 1

3 116.3

1 143.2

2 150% 142.5 145 3.91 97 2.73 149.6

DayAntibody Sample

Measured Potency

(%)

Mean Potency

(%)

SD (%)

Recovery (%)

CV (%)

1 38.4

2 50% 47.2 41.8 7.2 83.5 17.2

3 33.2

4 48.2

1 59.6

2 75% 70.2 67.4 5.2 89.9 7.7

3 69.3

4 70.5

1 120

2 125% 132.3 129.7 7.5 103.7 5.8

3 137.8

4 128.6

1 160.2

2 150% 158.2 162.8 5.2 108.5 3.2

3 162.7

4 170

Precision: 2.95%

Accuracy (recovery avg):

98.5%

Linearity: y = 0.922x + 5.0

Precision: 8.47%

Accuracy (recovery avg):

96.4%

Linearity: y = 1.22x - 21.3

Analyst 1: Analyst 2:

Linearity:



ADCC Reporter Bioassay (V Variant) is Specific

Assay signal is specifically

dependent on: Target cells expressing Ab-targeted

antigen

Specific antibody

Effector cells expressing FcgRIIIa

receptor



ADCC Reporter Bioassay is Robust

-10 -9 -8 -7 -6 -50

10

20

301

6

7

14


Fo

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f In

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-10 -9 -8 -7 -6 -50

10

20

30

1

1

7

13

16


Fo

ld o

f In

du

cti

on

Time of induction

E:T ratio and cell # per well

Run Induction time EC50

1 6.0hr 3.15x10-8 g/ml

2 5.5hr 3.83x10-8 g/ml

Run E:T ratio E cell # T cell # EC50

1 7.5:1 75k 10k 3.09x10-8 g/ml

2 6:1 90k 15k 3.83x10-8 g/ml



Stability Indicating for Fc Effector Function

EC50 = 5.77ng/ml

EC50 = 31.0ng/ml

Rituximab:

Tositumomab:

Trastuzumab:

Activity following heat-treatment of

antibody drugs



Miniaturizes to 384-Well Format

WIL2-S target cells Raji target cells

Assay volume per well Target cells Antibody Effector cells Bio-Glo™

96-well plate 25µl 25µl 25µl 75µl

384-well plate 5µl 5µl 5µl 15µl


March 2014

Antibody Variants, Glycosylation and Benchmarking with Classic ADCC Assay



Deglycosylated Herceptin

-12 -10 -8 -6 -40

5100 5

1100 6

2100 6

unt

10%unt/ 90% degly

EC50

unt

2.037e-008

10%unt/ 90% degly

7.174e-008

log [ab], (g/ml)

RL

U


-12 -10 -8 -6 -40

5100 5

1100 6

2100 6

unt

100% degly

EC50

unt

1.988e-008

100% degly

3.202e-008

log [ab], (g/ml)

RL

U


-12 -10 -8 -6 -40

5100 5

1100 6

2100 6

EC50

unt

1.720e-008

20%unt/80% degly

4.626e-008

unt

20%unt/80% degly

log [ab], (g/ml)

RL

U


-12 -10 -8 -6 -40

5100 5

1100 6

2100 6

unt

30%unt/ 70% degly

EC50

unt

2.002e-008

30%unt/ 70% degly

4.153e-008

log [ab], (g/ml)

RL

U


-12 -10 -8 -6 -40

5100 5

1100 6

2100 6

unt

40%unt/ 60% degly

EC50

unt

2.082e-008

40%unt/ 60% degly

3.486e-008

log [ab], (g/ml)R

LU


-12 -10 -8 -6 -40

5100 5

1100 6

2100 6

unt

50%unt/ 50% degly

EC50

unt

2.110e-008

50%unt/ 50% degly

2.990e-008

log [ab], (g/ml)

RL

U

Target cells: SKBR3; Unt = 100% glycosylated

Sensitive to Detect Differences in Glycosylation



Linear correlation obtained between

percentage of N-glycosylated antibody in

blended antibody samples and relative

luciferase reporter activity in ADCC

Reporter Bioassay

y = 0.0127x - 0.0314R² = 0.966

-0.100

0.000

0.100

0.200

0.300

0.400

0.500

0.600

0.700

0 10 20 30 40 50 60

Re

lati

ve a

ctiv

ity

in r

ep

ort

er

AD

CC

Percent N-glycosylated sample

y = 0.0125x - 0.0095R² = 0.9926

0.000

0.100

0.200

0.300

0.400

0.500

0.600

0.700

0 10 20 30 40 50 60

Re

lati

ve a

ctiv

ity

in r

ep

ort

er

AD

CC

Percent N-glycosylated sample

Rituximab

Trastuzumab

_____________________________________

Small differences in Fc effector activity in ADCC pathway activation are easily

distinguished in the ADCC Reporter Bioassay _____________________________________

Activity Correlates with Amount of Antibody N-Glycosylation



Sample set: Blended mixes of fully fucosylated and afucosylated Ab samples – provides a series with a range of

ADCC potencies. Relative activity: Expressed relative to 100% fucosylated Ab (lowest potency of series).

Both assays show increase in relative potency with increased % afucosylation of mAb

WIL2-S target cells; 6hr assay. E:T 6:1 LDH release assay; WIL2-S target cells; PBMCs

as effector cells. 4hr assay. E:T 25:1

Benchmarking Study: Good Correlation with Lytic LDH Release Assay

Classic ADCC assay(1) ADCC Reporter Bioassay(2)

1. Chung, S et al (2012). mAbs, 4(3), 326-340; doi: 10.4161/mabs.19941

2. Cheng, J et al (2012) AACR, poster #4606.


March 2014

Polymorphism in the FcgRIIIa Receptor

Optimizing the F Variant of the ADCC Reporter Bioassay



Effector:Target (E:T) Ratio

Suspension target cells Adherent target cells

10:1

6:1

4:1 15:1

10:1

ADCC Reporter Bioassay,

F Variant



Induction Time

Fresh-from-culture effector cells Frozen, thaw-and-use effector cells

24hr

7hr

5hr

6hr

24hr

Jurkat cell format ADCC Reporter Bioassay,

F Variant



Bioassay Development Using DOE

Factors:

Target cell density

Target cell/antibody

incubation time

Effector cell density

Induction time

Outputs:

Fold of induction

EC50

EC50 difference between F

and V variant Jurkat

effector cells

DOE = Design of Experiments

Allow understanding of the

interactions between critical

assay factors

Minimum amount of work

needed to develop robust

assays


F Variant



The F Variant Bioassay is Specific

Assay signal is specifically dependent on:

Target cells expressing Ab-targeted antigen

Specific antibody

Effector cells expressing FcgRIIIa receptor

No target cells

No Ab or wrong Ab

FcgRIIIa blocked

No FcgRIIIa



F Variant: Able to Measure Potencies of On-Market mAb Biologic Drugs

Rituximab, anti-CD20,

chimeric IgG1. Approved to

treat B-cell non-Hodgkin

lymphoma and chronic

lymphocytic leukemia.

Trastuzumab, anti-HER2,

humanized IgG1. Approved to

treat HER2-positive breast

cancer and stomach cancer.

Cetuximab, anti-EGFR,

chimeric IgG1. Approved to

treat colorectal cancer and

certain types of head and

neck cancer.

Target cell: SK-BR-3 Target cell: A431

Three best-selling, on-market mAb biologic drugs that posses ADCC as main MOA

Tests performed using ADCC Reporter Bioassay, Core Kit (F Variant)



Antibody IgG-Isotype Specificity

Order of response: hu IgG1, IgG3 > mouse

IgG2a >> hu IgG2, IgG4, mouse IgG1

Expanding assay applications:

1. Confirm desired ADCC activity for IgG1

and IgG3-based mAbs

2. Identify non-designed ADCC activity for

mAbs with non-ADCC MOA

2 1

Data generated using the ADCC Reporter

Bioassay, F Variant



Detects Loss of Activity Due to Heat-Stress

trastuzumab

EC50 increases (right shift)

Reporter bioassay exhibits stability-indicating capability

Can potentially be used in stability studies for therapeutic antibody

Fold induction

decreases


F Variant



Polymorphism in the FcgRIIIa Receptor Impacts ADCC Response

Because many new generation mAb drugs are being designed to improve Ab affinity for

the F variant of the receptor, drug developers need a low variability ADCC bioassay

specifically to measure the new drug potency via the F158 variant of FcgRIIIa.

Rituximab Trastuzumab

The dual receptor bioassay approach of the Promega ADCC Reporter Bioassay, with both V

and F variants of the FcgRIIIa allow researchers to better characterize their Ab drugs.


March 2014

Product Formats



Product Configurations: ADCC Reporter Bioassay, V Variant

Engineered Jurkat cells

Bio-Glo™ Reagent

Control Antibody

Target WIL2-S or Raji Cells

Core Kit

Complete

Kits

+ Medium Serum

1. Core Kits:

1X kit – Cat.# G7010

5X kit – Cat.# G7018

2. Complete Kits:

WIL2-S Target Cells – Cat.# G7014

Raji Target Cells – Cat.# G7015

3. Target Kits:

WIL2-S Target Cells – Cat.# G7013

Raji Target Cells – Cat.# G7016

4. Cell Propagation Model

ADCC Bioassay Effector Cells,

Propagation Model – Cat.# G7102

2 vials of Jurkat Effector cells:

allows propagation and banking for

use in ADCC via unique LULL

Target Kits

Multiple formats flexible to your

research needs:

Engineered

Jurkat cells,

V variant

Propagation

Model



Custom Product Configurations for ADCC Reporter Bioassay, F Variant

Engineered Jurkat

cells, F variant

Bio-Glo™

Reagent

Control

Antibody

Target WIL2-S or

Raji Cells

Core Kit

+ Medium Serum

Target Kits Catalog

products

Jurkat effector cells, F variant

Propagation

Model

1. Core Kit (Part# CS1324F01)

Components: ADCC Bioassay Effector Cells,

F Variant: in frozen, thaw-and-use

format

RPMI Medium

Low IgG Serum

Bio-Glo™ Luciferase Assay System

2. Cell Propagation Model

(Part# CS1324F08)

Components: ADCC Bioassay Effector Cells, F Variant,

Propagation Model: allows propagation

and banking for use in ADCC via unique

LULL with Bio-Glo™ Luciferase Assay

System



Ordering Information

ADCC Reporter Bioassays (V Variant)

Product Size Cat.

ADCC Reporter Bioassay, Core Kit 1ea G7010

ADCC Reporter Bioassay, Core Kit 5X 1ea G7018

ADCC Reporter Bioassay, Complete Kit (WIL2-S) 1ea G7014

ADCC Reporter Bioassay, Complete Kit (Raji) 1ea G7015

ADCC Reporter Bioassay, Target Kit (WIL2-S) 1ea G7013

ADCC Reporter Bioassay, Target Kit (Raji) 1ea G7016

ADCC Bioassay Effector Cells, Propagation Model 1ea Pls enquire

Contact: [email protected]

Bio-Glo™ Luciferase Assay System

100ml 10ml

G7940 G7941

Product Cat.

ADCC Reporter Bioassay, F Variant, Core Kit Pls enquire

ADCC Bioassay Effector Cells, F Variant, Propagation Model Pls enquire

Contact [email protected]

ADCC Reporter Bioassays (F Variant) Pre-commercial materials are available now through our

Custom Assay Services group. Simply e-mail CAS at the

address below.

mailto:[email protected]




Adopted by the Contract Services Industry

Approved manufacturing cell line switch by a pharmaceutical company

Submitted in an IND filing by a pharmaceutical company

In development for lot-release testing by a pharmaceutical company

Adopted by multiple pharmaceutical companies globally

BioAgilytix, Catalent, Covance & Charles River Laboratories are providing

ADCC Reporter Bioassay services



Highlights of the ADCC Reporter Bioassay…

Features Low variability

Engineered effector cells (Jurkat FcgRIIIa/NFAT-RE luc2)

replace primary NK cells

“Cells as reagents” - frozen, thaw-and-use format

Simple & robust protocol

Broad applicability in use with suspension or adherent

target cells

Good correlation with classic ADCC bioassay (cytolytic)

Benefits Demonstrates precision, accuracy, linearity, robustness,

specificity

Can quantify potency and stability of therapeutic Ab drugs

Can differentiate biological activity of Fc effector function

in ADCC MOA resulting from small changes in Ab

glycosylation


March 2014

Simple, Convenient Luminometer

Tested with ADCC Reporter Bioassay



Easy Reporter Assay Detection

GloMax® Discover System…

Integrated for Promega’s ADCC Reporter Bioassay

And ready to run any of the following reporter, cell

health and protein interaction assays

Product Size Cat.

GloMax® Discover Multimode Detection System 1ea GM3000

Cell Health:

CellTiter-Glo® Assay

CellTox™ Green Assay

Caspase-Glo® Assay

BacTiter-Glo® Assay

Luciferase Reporters:

Nano-Glo® Assay

ONE-Glo™ Assay

Dual-Glo® & DLR Assays

Bright-Glo™ Assay

Protein Interaction Assays:

ELISAs

BRET

FRET See www.promega.com/glomax for more

information

http://www.promega.com/glomax



GloMax® Discover System

Easy-To-Use

Choose from preloaded Promega

protocols or customize your own.

Build custom protocols using the intuitive

drag-and-drop interface.

Export data to a network, cloud or any

drive desired.

Superior Sensitivity

Plate mask (aperture control) for

switching between 96-well and 384-well

plates.

Low cross-talk between wells gives you

better, more usable results.

Broad dynamic range of the instrument

extends the linear range of your assay.

Minimal Manual Intervention

Filter paddles and automatic filter

switching allow easy two-color

multiplexing assays or kinetic studies.

Quick read-speeds for high efficiencies in

your laboratory.

Compatible for 3rd party automation

control.

Quality

IQ/OQ Service available.

Provides required technical

elements of a 21CFR 11 compliant

system to be used with the

appropriate laboratory workflow.



For More Information

Neal Cosby, PhD Strategic Marketing Manager [email protected] Custom Order Department [email protected] Or see: www.promega.com/adcc



http://www.promega.com/adcc

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