Development and validation of a cell-based fluorescent method for measuring antibody affinity.

ABSTRACT: Monoclonal antibodies have become essential tools for diagnostic and therapeutic purposes. Antibody affinity is one of the critical factors influencing the therapeutic success of tumor-targeting antibodies. Therefore, developing an accurate and reliable method for determining antibody affinity is crucial. In this study, we describe a fluorescent cell-based immunosorbent assay that can accurately measure antibody affinity (K_D) in the nanomolar range. This method involves the addition of fluorescently labeled antibodies to antigen-positive and antigen-negative cell lines fixed on 96-well plates. The fluorescent signals from nonspecific binding to negative control cell lines is subtracted from the specific binding to the antigen-positive cell lines. The K_D values obtaine

SUBMITTER: Yu X 

PROVIDER: S-EPMC5293658 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature