2004•Chinese journal of biotechnology/Shengwu gongcheng xuebaoRequires access
Study on the Methods of Purification of Recombinant Humanized anti-HBsAg Fab Antibody
Ning Deng, Junjian Xiang, Rao G, Chen Wenyin
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Abstract
Anti-HBsAg Fab antibody is useful in prevention if it binds to and block surface regions of the virus, which is essential for the infection of hepatocytes. In order to establish steady and high effective methods to purify the recombinant humanized anti-HBsAg Fab antibody from fermentation supernatant of Pichia pastoris for industrial production,anti-Fab chromatography, anti-ScFv Mab chromatography and ion exchange chromatography were used. Results showed that the purity of recombinant anti-HBsAg Fab antibody purified by affinity chromatography of anti-Fab antibody column is 96.8%, and the recovery rate of the Fab antibody is about 30%～40% and much of Fab fragment was lost. The purity of recombinant anti-HBsAg Fab antibody purified by affinity chromatography of anti-ScFv Mab column is 97.5% and the recovery rate of the Fab antibody is about 75%～85%. This method can be used in smaller scale purification of recombinant Fab antibody from fermentation supernatant of recombinant yeast. The purity of recombinant Fab fragment purified by ion exchange chromatography is 97%, and the recovery rate of the Fab antibody is about 75%～85%. The method of ion exchange chromatography can be used to produce the recombinant Fab antibody in large scale for industrial production. The results demonstrated that the recombinant humanized anti-HBsAg Fab antibody can be purified high effectively by the methods of anti-ScFv Mab chromatography and ion exchange chromatography from fermentation supernatant of recombinant yeast, and the methods of anti-ScFv Mab chromatography and ion exchange chromatography can be used effectively in industrial production.