PURIFICATION OF SOLUBLE RECOMBINANT HUMAN FcγRII (CD32)

Abstract

The present study describes the methodology used to purify human recombinant low-affinity FcγRIIa₂ 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-FcγRIIa_1/2 immunosorbent, followed by gel filtration chromatography. Using this method, the purified recombinant FcγRIIa₂ 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 FcγRIIa₂ for further characterization of its biological activities.

^*Work for which these authors equally contributed.

ACKNOWLEDGMENTS

The authors wish to thank Mr. Lucien Cabanié (Institut Curie, Paris, France) for his expert technical assistance. This work was supported by the Institut Curie, the INSERM, and the Association pour la Recherche contre le Cancer (ARC). JC was supported by a fellowship from the Fondation pour la Recherche Médicale.

Notes

^*Work for which these authors equally contributed.