title: Membrane-based receptor affinity chromatography.
abstract: Membrane-based receptor affinity chromatography (MRAC), which utilizes the molecular recognition between an immobilized receptor and its soluble protein ligand, has been developed for the purification of human interleukin-2 and related biomolecules. The multi-purpose affinity membrane used in this study consisted of a soluble form of interleukin-2 receptor (IL-2R) chemically bonded to hollow-fiber membranes in an oriented fashion. A model system involving anti-Tac-H (a humanized monoclonal antibody to IL-2R) was used to study the important factors influencing the performance of MRAC, including support morphology, mass transfer rate and adsorption kinetics. All three are shown to be highly efficient. MRAC has been successfully applied to the purification of anti-Tac-H, recombinant human interleukin-2 (rIL-2) and interleukin 2-Pseudomonas exotoxin fusion protein (IL2-PE40). Overall, MRAC was found to be a viable, scalable and extremely productive affinity purification method.
authors.affiliation: Protein Biochemistry Department, Roche Research Center, Hoffmann-La Roche Inc., Nutley, NJ 07110.
authors.forename: M
authors.initials: M
authors.lastname: Nachman
authors.forename: A R
authors.initials: AR
authors.lastname: Azad
authors.forename: P
authors.initials: P
authors.lastname: Bailon
mesh_terms: D000327:Adsorption; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000302:isolation & purification; D001665:Binding Sites; D002846:Chromatography, Affinity; D004058:Diffusion; D005098:Exotoxins; D000922:Immunotoxins; D007376:Interleukin-2 / Q000276:immunology / Q000302:isolation & purification*; D008024:Ligands; D008566:Membranes / Q000737:chemistry; D010084:Oxidation-Reduction; D011955:Receptors, Drug / Q000737:chemistry*; D011994:Recombinant Proteins / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D005098:Exotoxins; C057961:IL-2-PE40 chimeric protein, recombinant; D000922:Immunotoxins; D007376:Interleukin-2; D008024:Ligands; D011955:Receptors, Drug; D011994:Recombinant Proteins