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NEUTRALIZING ANTIBODIES MAKING THEIR MARK IN NEXT WAVE OF BIOLOGICS

Antibody Internalization: Advanced Flow
Cytometry and Live-Cell Analysis Give Rich
Insights During Antibody Profiling
CLARE SZYBUT1*, CAROLINE WELDON2*, NICOLA BEVAN1*, LORI KING
1.	 1. Essen BioScience, Ltd., Part of the Sartorius Group, Units 2 & 3 The Quadrant, Newark Close, Royston Hertfordshire SG8 5HL, UK
2.	 2. Essen BioScience, Inc., Part of the Sartorius Group, 5700 Pasadena Ave. NE, Albuquerque, NM 87113 USA

Introduction

T

candidates. For instance, binding and rapid

he natural characteristics of antibodies,

internalization are desirable properties for

such as high binding affinity, specificity

antibody-drug conjugates (ADCs), as cells must

to a wide variety of targets, and

be selectively targeted and killed via delivery

good stability, make them ideal therapeutic

of a cytotoxic payload. In contrast, if the goal is

candidates for many diseases. Monoclonal

to induce antibody-dependent cell-mediated

antibodies (mAbs), in particular, deliver promising

cytotoxicity (ADCC), the antibody must remain

therapeutic results in several different disease

bound to the cell surface, rather than being

areas, such as autoimmunity, oncology, and

internalized, in order to activate an immune

chronic inflammation. Researchers' abilities to

response.

improve the breadth of antibodies have been

When designing ADCs, one key attribute to

aided by innovative technologies for antibody

predicting efficacy is the antibody internalization

discovery, for instance, through humanization of

(ABI) rate and the associated kinetics. These

mouse antibodies and phage display. However,

internalization kinetics are influenced by factors

advanced antibody design techniques create

such as the epitope on the target antigen, affinity

the need for new screening methods so that

of the ADC-antigen interaction, and intracellular

lead candidates can be quickly and effectively

trafficking. Evaluating for these factors is critical

identified as early in the development process as

throughout the antibody screening pathway

possible.

(Figure 1); however, here, we concentrate on

Part of an effective screening strategy is to
identify the desired therapeutic goal for your
8|

GENengnews.com

evaluating antibodies' ABI during functional
profiling via rate comparisons during the


http://www.GENengnews.com

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