Characterization of monoclonal antibody's binding kinetics using oblique-incidence reflectivity difference approach.

Abstract:

:Monoclonal antibodies (mAbs) against human proteins are the primary protein capture reagents for basic research, diagnosis, and molecular therapeutics. The 2 most important attributes of mAbs used in all of these applications are their specificity and avidity. While specificity of a mAb raised against a human protein can be readily defined based on its binding profile on a human proteome microarray, it has been a challenge to determine avidity values for mAbs in a high-throughput and cost-effective fashion. To undertake this challenge, we employed the oblique-incidence reflectivity difference (OIRD) platform to characterize mAbs in a protein microarray format. We first systematically determined the Kon and Koff values of 50 mAbs measured with the OIRD method and deduced the avidity values. Second, we established a multiplexed approach that simultaneously measured avidity values of a mixture of 9 mono-specific mAbs that do not cross-react to the antigens. Third, we demonstrated that avidity values of a group of mAbs could be sequentially determined using a flow-cell device. Finally, we implemented a sequential competition assay that allowed us to bin multiple mAbs that recognize the same antigens. Our study demonstrated that OIRD offers a high-throughput and cost-effective platform for characterization of the binding kinetics of mAbs.

journal_name

MAbs

journal_title

mAbs

authors

Liu S,Zhang H,Dai J,Hu S,Pino I,Eichinger DJ,Lyu H,Zhu H

doi

10.4161/19420862.2014.985919

subject

Has Abstract

pub_date

2015-01-01 00:00:00

pages

110-9

issue

1

eissn

1942-0862

issn

1942-0870

journal_volume

7

pub_type

杂志文章

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