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Anal Biochem. 2016 Sep 1;508:78-96. doi: 10.1016/j.ab.2016.06.024. Epub 2016 Jun 27.

Comparison of biosensor platforms in the evaluation of high affinity antibody-antigen binding kinetics.

Author information

1
Department of Immune Modulation and Biotherapeutics Discovery, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT 06877, USA.
2
Department of Immune Modulation and Biotherapeutics Discovery, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT 06877, USA; The Fu Foundation School of Engineering and Applied Science, Columbia University, New York, USA.
3
Department of Immune Modulation and Biotherapeutics Discovery, Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT 06877, USA. Electronic address: rachel.kroe-barrett@boehringer-ingelheim.com.

Abstract

The acquisition of reliable kinetic parameters for the characterization of biomolecular interactions is an important component of the drug discovery and development process. While several benchmark studies have explored the variability of kinetic rate constants obtained from multiple laboratories and biosensors, a direct comparison of these instruments' performance has not been undertaken, and systematic factors contributing to data variability from these systems have not been discussed. To address these questions, a panel of ten high-affinity monoclonal antibodies was simultaneously evaluated for their binding kinetics against the same antigen on four biosensor platforms: GE Healthcare's Biacore T100, Bio-Rad's ProteOn XPR36, ForteBio's Octet RED384, and Wasatch Microfluidics's IBIS MX96. We compared the strengths and weaknesses of these systems and found that despite certain inherent systematic limitations in instrumentation, the rank orders of both the association and dissociation rate constants were highly correlated between these instruments. Our results also revealed a trade-off between data reliability and sample throughput. Biacore T100, followed by ProteOn XPR36, exhibited excellent data quality and consistency, whereas Octet RED384 and IBIS MX96 demonstrated high flexibility and throughput with compromises in data accuracy and reproducibility. Our results support the need for a "fit-for-purpose" approach in instrument selection for biosensor studies.

KEYWORDS:

Antibody-antigen interactions; Biacore; MX96; Octet; Optical biosensor; ProteOn

PMID:
27365220
DOI:
10.1016/j.ab.2016.06.024
[Indexed for MEDLINE]
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