[The property of protein A affinity membrane cartridge and purification of IgG from human plasma].
A new protein A affinity membrane based on composite matrix formed by covalent cross-linking of the glycidyl methylacrylate to softwood cellulose fiber was developed. The study focused on the physical properties and adsorption capacity of the affinity membrane cartridge. The results showed that the relationship between flow-rate and backpressure was linear. When flow-rate was 3 mL/min, backpressure attained 160 kPa. The effects of IgG mass concentration and sampling flow-rate on the binding capacity by protein A cartridge were studied for the optimum purification of IgG. A 21.7 mg/g dry matrix of maximum dynamic binding capacity for IgG was obtained by dynamic adsorption. The cartridge was loaded with human plasma to test its ability to fractionate IgG. The IgG purified from human plasma was analysed by capillary zone electrophoresis (CZE). The results showed that the protein A affinity membrane cartridge had high selectivity for IgG from human plasma. The purity of IgG from human plasma purified by the immobilized protein A membrane cartridge was over 98%. The fast-flow property of the membrane cartridge made from such a matrix and its simplicity in operation provide effective means for purifying immunoglobulins on a relatively large scale.
Jia
L
L
National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116012.
Yang
L
L
Zou
H
H
Zhang
Y
Y
D002846:Chromatography, Affinity / Q000379:methods; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification*; D008567:Membranes, Artificial; D010949:Plasma / Q000276:immunology*; D013205:Staphylococcal Protein A
D007074:Immunoglobulin G; D008567:Membranes, Artificial; D013205:Staphylococcal Protein A