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Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms

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Using multiple platforms for critical reagents selection process to support pharmacokinetic ligand-binding assay

Elisa Oquendo¹, Xu Lin¹, Sally Ye¹

¹Boehringer Ingelheim Pharmaceuticals Inc., Drug Metabolism & Pharmacokinetics, Ridgefield, CT 06877, USA.

|March 26, 2021

Summary

Biolayer interferometry (BLI) epitope binning showed limited success in predicting antibody pairs for pharmacokinetic assays. While BLI identified potential pairs, many did not perform well in subsequent ELISA and Gyrolab assays.

Keywords:

antibody pairing biolayer interferometry ligand binding assays pharmacokinetic assays pharmacokinetic pairing

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Area of Science:

Biochemistry
Immunology
Analytical Chemistry

Background:

Pharmacokinetic (PK) assay development requires reliable antibody pairs.
Epitope binning is a technique used to characterize antibody interactions.
Biolayer interferometry (BLI) is a label-free method for analyzing biomolecular interactions.

Purpose of the Study:

To evaluate the utility of epitope binning using BLI for selecting candidate antibody pairs for PK assay development.
To compare the predictive power of BLI epitope binning against ELISA and Gyrolab assays.
To assess the correlation between BLI results and successful antibody pair performance in downstream assays.

Main Methods:

Epitope binning was performed using BLI with two formats: in-tandem and classic sandwich.
Eighty anti-idiotypic monoclonal antibodies were tested in 64 combinations.
Results were compared with those obtained from ELISA and Gyrolab assays.

Main Results:

BLI identified four mutually exclusive epitope bins, yielding 31 (in-tandem) and 25 (classic sandwich) antibody pair combinations.
ELISA identified 18 positive pairs, and Gyrolab identified 9 positive pairs.
A partial correlation was observed between BLI and the other assays; several pairs positive by ELISA/Gyrolab were negative by BLI, and only 50% of BLI-predicted pairs were positive by ELISA, with less than 25% positive by Gyrolab.

Conclusions:

BLI epitope binning demonstrated limitations in predicting successful antibody pairs for PK assay development in this study.
The predictive accuracy of BLI for downstream assay performance was found to be suboptimal.
Further optimization or alternative methods may be needed for efficient candidate antibody pair selection in PK assay development.