Simple multiplexed antibody-binding assay

Abstract

This simple protocol tests antibody binding to target antigens on the surface of cells. This assay is powerful because negative controls are built into each well of the screening plates. It can be used to screen crude supernatants from hybridomas, as well as bacterial periplasmic extracts when screening phage libraries. Using cell-permeant dyes allows the negative and positive cell lines to be color-coded and screened in the same well. A variant enzyme-linked immunosorbent assay can be performed where the target antigen is presented on beads.