title: Characterization of labeled reagents in ligand-binding assays by a surface plasmon resonance biosensor.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.affiliation: BioAnalytical Sciences, Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Jia
authors.initials: J
authors.lastname: Duo
authors.affiliation: Sierra Sensors GmbH, Hamburg 20251, Germany.
authors.forename: JoAnne
authors.initials: J
authors.lastname: Bruno
authors.affiliation: BioAnalytical Sciences, Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Steven
authors.initials: S
authors.lastname: Piccoli
authors.affiliation: BioAnalytical Sciences, Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Binodh
authors.initials: B
authors.lastname: DeSilva
authors.affiliation: BioAnalytical Sciences, Analytical & Bioanalytical Operations, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Yan J
authors.initials: YJ
authors.lastname: Zhang
mesh_terms: D061067:Antibodies, Monoclonal, Humanized / Q000737:chemistry / Q000378:metabolism; D015374:Biosensing Techniques; D001710:Biotin / Q000737:chemistry; D019921:Biotinylation; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D008163:Luminescent Measurements; D011092:Polyethylene Glycols / Q000737:chemistry; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D020349:Surface Plasmon Resonance
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D055767:Immobilized Proteins; D008024:Ligands; D011506:Proteins; D011092:Polyethylene Glycols; D001710:Biotin
keywords: binding affinity; binding kinetics; critical reagents; ligand-binding assay; reagent labeling; surface plasmon resonance (SPR) biosensor