Sartorius_May21_NeutAntiMakingTheir - 9

NEUTRALIZING ANTIBODIES MAKING THEIR MARK IN NEXT WAVE OF BIOLOGICS

Ab raised as
multiple hybridomas
(1000s Ab)

Early cellular
testing for
functionality,
affinity ranking

Check for binding
to target (500-1000
Ab)

Single
concentration
screening of 100s
Ab, non-purified,
direct comparison

Advanced HighThroughput
Flow Cytometry

Kinetic LiveCell Analysis

Scale up of key Ab
(~50)

Humanization
of Ab of interest
(<10)

Further functional
evaluation

Full concentration
range profiling

Full concentration
range profiling, rate
comparison

Full concentration
range profiling,
mechanistic studies
multiplexed with
additional readouts

Figure 1: Inclusion of ABI in the screening pathway for antibodies. Selecting for ABI throughout the antibody screening pathway,
whether during functional profiling and rate comparisons or mechanistic studies.

development process, as well as multiplexed

insight by using a novel, pH-sensitive reagent

mechanistic studies later in the screening process.

for characterizing ABI via both advanced flow

Traditionally, antibody screening

cytometry and live-cell imaging.

workflows have required labor intensive, time
consuming, end-point-only methods, such

Challenges with Traditional

as FACS, ELISA, or microscopy. One solution

Antibody Screening Workflows

to address these screening challenges is to

Monoclonal antibodies (mAbs) are large

use advanced screening methods that offer

(about 150 kDa), complex biologic molecules

maximum insight through high-content data.

that require post-translational modifications

Herein, we demonstrate an advanced antibody

for their activity (Chames et al., 2009).

screening method that focuses on speed and

Therefore, researchers face several challenges

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Sartorius_May21_NeutAntiMakingTheir

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