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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.
Supplementary Data
Financial & competing interests disclosure
All financial support for this work was received from the Bristol-Myers Squibb Company (NJ, USA). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.
No writing assistance was utilized in the production of this manuscript.
Ethical conduct of research
The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.
Acknowledgements
The authors sincerely thank H Myler for her scientific review and valuable input to the manuscript.