Unit

UNIT 18.2 Detection of Low-Affinity Ligand-Receptor Interactions at the Cell Surface with Fluorescent Microspheres

  1. Marion H. Brown

Published Online: 1 MAY 2002

DOI: 10.1002/0471142735.im1802s48

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Brown, M. H. 2002. Detection of Low-Affinity Ligand-Receptor Interactions at the Cell Surface with Fluorescent Microspheres. Current Protocols in Immunology. 48:18.2:18.2.1–18.2.6.

Author Information

  1. Medical Research Council Cellular Immunology Unit, Sir William Dunn School of Pathology, Oxford, United Kingdom

Publication History

  1. Published Online: 1 MAY 2002
  2. Published Print: APR 2002

Abstract

A method of producing highly avid multivalent ligand binding reagents for detecting low-affinity interactions at the cell surface is described in this unit. The principle is to immobilize multiple copies of extracellular regions of cell-surface molecules on plastic fluorescent beads, to present the coated beads to cells, and to analyze binding in a quantitative manner by flow cytometry. The method of attaching proteins to the beads is designed to maximize display of the ligand-binding region. The approach is applicable to immobilization of fusion proteins on a variety of beads, and can also be adapted for use with native proteins. A protocol for the biotinylation of MAbs is also presented.