Centrifugation assay of IgE‐mediated cell adhesion to antigen‐coated gels

Abstract

AbstractThe adhesion of biological cells to substrates is often mediated by binding between cellular receptors and substrate‐bound ligand. In this work, we used a centrifugation assay to measure the adhesion of rat basophilic leukemia (RBL) cells coated with immunoglobulin E (IgE) to substrates coated with the ligand dinitrophenol (DNP). Increasing force, decreasing DNP substrate density, and decreasing cell surface IgE density all led to decreasing adhesion. Experiments performed at low IgE cell surface densities, in which few tethers from between cell and substrate suggest individual tethers have a binding strength of 2 to 4 microdyne, in agreement with previous measurements of the force to uproot receptors from the plasma membrane. We use this system to show how subpopulations expressing different numbers of cell surface receptors may be separated by exploiting their differential adhesiveness to substrates.