Determination of equilibrium dissociation constants for recombinant antibodies by high-throughput affinity electrophoresis.

ABSTRACT: High-quality immunoreagents enhance the performance and reproducibility of immunoassays and, in turn, the quality of both biological and clinical measurements. High quality recombinant immunoreagents are generated using antibody-phage display. One metric of antibody quality - the binding affinity - is quantified through the dissociation constant (K_D) of each recombinant antibody and the target antigen. To characterize the K_D of recombinant antibodies and target antigen, we introduce affinity electrophoretic mobility shift assays (EMSAs) in a high-throughput format suitable for small volume samples. A microfluidic card comprised of free-standing polyacrylamide gel (fsPAG) separation lanes supports 384 concurrent EMSAs in 30 s using a single power source. Sample is disp

SUBMITTER: Pan Y 

PROVIDER: S-EPMC5180089 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature