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28 Sep 2019
An antibody is found to bind an antigen of interest with association rate constant ka = 2x104M-1s-1 and dissociation rate constant kd = 6x10-4s-1. In an attempt to affinity mature this antibody, a set of CDR point mutants were generated and screened by an ELISA binding assay, using a single concentration of 300nM of each antibody variant. Very little improvement in the amount of binding was observed among the variants (at best 10% more than the parental antibody). However, additional analysis of the binding kinetics showed that some of these variants have as much as 100 times higher affinity. Why were these differences not observed in the primary ELISA screen?
An antibody is found to bind an antigen of interest with association rate constant ka = 2x104M-1s-1 and dissociation rate constant kd = 6x10-4s-1. In an attempt to affinity mature this antibody, a set of CDR point mutants were generated and screened by an ELISA binding assay, using a single concentration of 300nM of each antibody variant. Very little improvement in the amount of binding was observed among the variants (at best 10% more than the parental antibody). However, additional analysis of the binding kinetics showed that some of these variants have as much as 100 times higher affinity. Why were these differences not observed in the primary ELISA screen?
Collen VonLv2
28 Sep 2019
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