Jia Duo1, JoAnne Bruno2, Steven Piccoli1, Binodh DeSilva1, Yan J Zhang1. 1. BioAnalytical Sciences, Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA. 2. Sierra Sensors GmbH, Hamburg 20251, Germany.
Abstract
AIM: Ligand-binding assay (LBA) reagent labeling may change the binding characteristics of the reagent to its target and degrade its performance in LBAs. RESULTS: A surface plasmon resonance (SPR) biosensor was used to evaluate the impact of the biotin labeling process on reagent-binding kinetics and affinity for a specific target. The SPR results demonstrate that the biotin molar challenge ratio affects both association and dissociation rates for the labeled reagent binding to its target. The SPR results also predict the labeled reagent performance in LBAs. CONCLUSION: The methodology used in this study provides an example of using an SPR biosensor as an efficient way to analytically and functionally characterize critical reagents and to understand their performance postmodification in LBAs.
AIM: Ligand-binding assay (LBA) reagent labeling may change the binding characteristics of the reagent to its target and degrade its performance in LBAs. RESULTS: A surface plasmon resonance (SPR) biosensor was used to evaluate the impact of the biotin labeling process on reagent-binding kinetics and affinity for a specific target. The SPR results demonstrate that the biotin molar challenge ratio affects both association and dissociation rates for the labeled reagent binding to its target. The SPR results also predict the labeled reagent performance in LBAs. CONCLUSION: The methodology used in this study provides an example of using an SPR biosensor as an efficient way to analytically and functionally characterize critical reagents and to understand their performance postmodification in LBAs.
Authors: Kun Yang; Ying Zhang; Robert Chou; Lai Yeung; Simon Letarte; Rong-Sheng Yang; Xuanwen Li; Maribel Beaumont; Rico Gunawan; Douglas Richardson; Shara Dellatore; Eric Woolf; Yang Xu Journal: ACS Pharmacol Transl Sci Date: 2020-10-28