title: Affinity purification of antibodies using immobilized FB domain of protein A.
abstract: A continuous method for the efficient digestion of protein A into active fragments (FB, Mr = 7000) using immobilized trypsin was developed. These fragments originate from almost identical five-repeated monovalent Fc-binding units of 58 residues each. The fragments obtained were found to be similar to the recently described genetically engineered fragment B. Antibody-binding characteristics of the FB domain and also of intact protein A, immobilized on to adipic dihydrazide-modified Eupergit CB6200 beads, were investigated. Based on the experimental data obtained, a high-performance liquid chromatographic column containing C30N Eupergit C-immobilized FB domain was prepared and its performance in antibody purification was compared with that of Eupergit C-immobilized intact protein A.
authors.affiliation: Department of Molecular Microbiology and Biotechnology, George Wise Faculty of Life Sciences, Tel-Aviv University, Israel.
authors.forename: B
authors.initials: B
authors.lastname: Solomon
authors.forename: O
authors.initials: O
authors.lastname: Raviv
authors.forename: E
authors.initials: E
authors.lastname: Leibman
authors.forename: G
authors.initials: G
authors.lastname: Fleminger
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000906:Antibodies / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000276:immunology; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D016650:Fluorescein-5-isothiocyanate; D006868:Hydrolysis; D010446:Peptide Fragments / Q000302:isolation & purification*; D013205:Staphylococcal Protein A / Q000276:immunology / Q000302:isolation & purification*; D014357:Trypsin
chemical_list: D000596:Amino Acids; D000906:Antibodies; D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D013205:Staphylococcal Protein A; D014357:Trypsin; D016650:Fluorescein-5-isothiocyanate