Unit

UNIT 2.9 Purification of Immunoglobulin M and Immunoglobulin D

  1. Sarah M. Andrew (IgM)1,
  2. Julie A. Titus (IgM)2,
  3. Richard Coico (IgD)3,
  4. Ashok Amin (IgD)4

Published Online: 1 MAY 2001

DOI: 10.1002/0471142735.im0209s21

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Andrew, S. M., Titus, J. A., Coico, R. and Amin, A. 2001. Purification of Immunoglobulin M and Immunoglobulin D. Current Protocols in Immunology. 21:III:2.9:2.9.1–2.9.8.

Author Information

  1. 1

    Lancaster University, Lancaster, United Kingdom

  2. 2

    National Cancer Institute, Bethesda, Maryland

  3. 3

    City University of New York Medical School, New York, New York

  4. 4

    Hospital for Joint Diseases, New York, New York

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: MAR 1997

This is not the most recent version of the article. View current version (1 APR 2009)

Abstract

This unit describes two classical protocols for the purification of IgM - dialysis of ascites fluid, tissue culture medium, or bioreactor supernatants against distilled water to precipitate pure IgM, and ammonium sulfate precipitation. Both protocols can be followed by size-exclusion chromatography to obtain a highly purified product. Recently an affinity method for purification of IgM has been developed using mannan binding protein, and is described here. The third approach presented is a one-step IgD purification method, designed specifically for murine derived samples, that uses Sepharose coupled to lectin derived from the seeds of Griffonia simplicifolia-1. This represents a simple, rapid, and gentle approach to isolating this highly labile immunoglobulin from IgD-containing ascites or hybridoma sources.