title: IgG purification using affinity filtration with sulfamethazine-affinity carriers.
abstract: Immunoglobulin G (IgG) antibodies are used extensively for analytical, diagnostic, and therapeutic applications. However, there are some disadvantages to purify IgG antibodies by protein A and G affinity chromatography. Therefore, it is necessary to find an effective alternative and nonchromatographic method to purify IgG. Dextran microparticles were activated and coupled with sulfamethazine to form sulfamethazine-affinity carriers. Then the carriers were used to purify IgG by affinity filtration. Quantitative and qualitative determination proved that sulfamethazine would successfully bond to the surface of dextran microparticles with a density of 85.5 μmol/g (wet). Affinity carriers were proved to withstand high shear force and reveal rare sulfamethazine leakage under filtration conditions between pH 3 to 11. The maximum IgG-binding capacity of affinity carriers was 8.03 mg IgG/g (wet). The affinity filtration process obtained a recovery yield above 80% and purity above 90%. Thus, this work involved in both the advantages of membrane filtration and affinity purification. The results, for the first time, proved that it is possible to use the small ligand sulfamethazine for affinity filtration of IgG. It is an attractive alternative to conventional protein A or G affinity chromatography.
authors.affiliation: College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Yi
authors.forename: Li
authors.initials: L
authors.lastname: Zhu
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Mei
authors.forename: Jianshu
authors.initials: J
authors.lastname: Chen
authors.forename: Guoqing
authors.initials: G
authors.lastname: Ying
mesh_terms: D000327:Adsorption; D000818:Animals; D000915:Antibody Affinity; D001201:Ascites / Q000276:immunology; D002482:Cellulose / Q000031:analogs & derivatives / Q000737:chemistry; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D003911:Dextrans / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004811:Epichlorohydrin / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification*; D008024:Ligands; D008567:Membranes, Artificial; D051379:Mice; D011485:Protein Binding; D013418:Sulfamethazine / Q000737:chemistry*
chemical_list: D003911:Dextrans; D007074:Immunoglobulin G; D008024:Ligands; D008567:Membranes, Artificial; D004811:Epichlorohydrin; C005062:acetylcellulose; D013418:Sulfamethazine; D002482:Cellulose