Purification of soluble recombinant human FcgammaRII (CD32)

Prep Biochem Biotechnol. 2001 Nov;31(4):341-54. doi: 10.1081/PB-100107481.

Abstract

The present study describes the methodology used to purify human recombinant low-affinity FcgammaRIIa2 produced in E. coli and to evaluate its binding to surface IgG. The recombinant molecule was purified by a two-step chromatographic procedure, including affinity chromatography using IV.3 anti-FcgammaRIIa1/2 immunosorbent, followed by gel filtration chromatography. Using this method, the purified recombinant FcgammaRIIa2 was 99% pure. It exhibited an isoeletric point of 5.2. Binding studies demonstrated a specific binding of the purified recombinant molecule to surface IgG expressed by human B cells. Thus, we have set up a method which allows to purify functional human recombinant FcgammaRIIa2 for further characterization of its biological activities.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • B-Lymphocytes / immunology
  • Binding Sites, Antibody / immunology
  • Chromatography, Affinity / methods*
  • Chromatography, Gel / methods*
  • Escherichia coli / genetics*
  • Humans
  • Immunoglobulin G / immunology
  • Receptors, IgG / immunology
  • Receptors, IgG / isolation & purification*
  • Recombinant Proteins / isolation & purification
  • Solubility

Substances

  • Immunoglobulin G
  • Receptors, IgG
  • Recombinant Proteins