Unit

UNIT 8.2 Immunoaffinity Chromatography

  1. Timothy A. Springer

Published Online: 1 MAY 2001

DOI: 10.1002/0471142735.im0802s18

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Springer, T. A. 2001. Immunoaffinity Chromatography. Current Protocols in Immunology. 18:II:8.2:8.2.1–8.2.9.

Author Information

  1. Center for Blood Research, Harvard Medical School, Boston, Massachussets

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUN 1996

Abstract

This unit describes the isolation of soluble or membrane-bound protein antigens from cells or homogenized tissue by immunoaffinity chromatography. The technique involves the elution of a single protein from an immunoaffinity column after prior elution of nonspecifically adsorbed proteins. To elute the bound antigen from the immunoaffinity matrix, the antibody-antigen interaction is destabilized by brief exposure to high-pH or low-pH buffer. The use of batch purification of antigens is an alternate procedure and results in shorter column loading times. The detergent octyl β-D-glucoside can be used instead of Triton X-100 for elution. Because octyl β-D-glucoside has a high critical micelle concentration (CMC), it can be readily removed by dialysis, as described.