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Binding sites for Ag

  • mtarca
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6 years 4 months ago #1 by mtarca
Binding sites for Ag was created by mtarca
Dear SPR,
How do I determine if my analyte has 2 binding sites. I noticed on a recent Biacore experiment that the experimental Rmax is approximately 50% higher than the calculated theoretical.

Thanks.

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  • Arnoud
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6 years 4 months ago #2 by Arnoud
Replied by Arnoud on topic Binding sites for Ag
Hi,

These two papers describe some methods you can use.
1. Day, E. S., Capili, A. D., Borysenko, C. W., et al.; Determining the affinity and stoichiometry of interactions between unmodified proteins in solution using Biacore. Analytical Biochemistry (440) 1: 96-107; 2013.
2. Oda, M. and Azuma, T.; Reevaluation of stoichiometry and affinity/avidity in interactions between anti-hapten antibodies and. Molecular Immunology (37) 18: 1111-1122; 2000.

Alternative methods to investigate the stoichiometry of an interaction are gel filtration (when the binding component add sufficient mass to be separated on the column), iso thermal calorimetry, analytical ultracentrifugation and dynamic light scattering.

Kind regards
Arnoud

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