title: Purification of soluble recombinant human FcgammaRII (CD32).
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.
authors.affiliation: Laboratoire de Biotechnologie des Anticorps, Institut Curie, Paris, France.
authors.forename: N
authors.initials: N
authors.lastname: Gruel
authors.forename: J
authors.initials: J
authors.lastname: Chapiro
authors.forename: W H
authors.initials: WH
authors.lastname: Fridman
authors.forename: J L
authors.initials: JL
authors.lastname: Teillaud
mesh_terms: D001402:B-Lymphocytes / Q000276:immunology; D001666:Binding Sites, Antibody / Q000276:immunology; D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D017452:Receptors, IgG / Q000276:immunology / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification; D012995:Solubility
chemical_list: D007074:Immunoglobulin G; D017452:Receptors, IgG; D011994:Recombinant Proteins