Surface Plasmon Resonance (SPR) as a Tool for Antibody Conjugate Analysis

Surface plasmon resonance (SPR) analysis was used to assess the immunoreactivity of anti-biotin (4) and anti-fluorescein (5) monoclonal antibody after conjugation with the N-hydroxysuccinimide ester of acridinium-9-carboxamide 1. Only minor changes in the apparent equilibrium dissociation constants...

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Bibliographic Details
Published in:Bioconjugate chemistry 1999-11, Vol.10 (6), p.1032-1037
Main Authors: Adamczyk, Maciej, Mattingly, Phillip G, Shreder, Kevin, Yu, Zhiguang
Format: Article
Language:English
Subjects:
Acridines - chemistry
Antibodies, Monoclonal - analysis
Antibodies, Monoclonal - chemistry
Binding, Competitive
Biosensing Techniques
Biotin - immunology
Fluorescein - chemistry
Solutions
Surface Plasmon Resonance
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Summary:Surface plasmon resonance (SPR) analysis was used to assess the immunoreactivity of anti-biotin (4) and anti-fluorescein (5) monoclonal antibody after conjugation with the N-hydroxysuccinimide ester of acridinium-9-carboxamide 1. Only minor changes in the apparent equilibrium dissociation constants of the antibody conjugates for their ligands resulted from the conjugation process. However, comparison of the initial binding rate of the conjugates with their ligands with those of the unmodified antibodies over a range of concentrations showed that the antibody conjugates were partially inactivated. The anti-fluorescein conjugates retained at least 90% of their immunoreactivity over the range of modification tested, while anti-biotin conjugates showed a progressive loss of reactivity with increased substitution by the label.
ISSN:1043-1802
1520-4812
DOI:10.1021/bc990057e