人源性抗HBsAg抗体Fab段在酵母中的表达

The Expression of Humanized Fab Fragment of the Anti-HBsAg Antibody in Methylotropic Yeast Pichia pastoris

通过分步整合的方式 ,将人源性抗乙肝表面抗原 (HBsAg)抗体Fab的轻、重链基因分步整合到巴斯德毕赤(Pichiapastoris)酵母GS115菌株的染色体上 ,经甲醇诱导 ,成功地分泌表达出抗HBsAg抗体的Fab片段 ,表达量达5 0～ 80mg L。ELISA结果显示重组酵母分泌表达出的Fab具有较强的结合HBsAg的能力。通过抗Fab的抗体柱亲和层析 。

Using of two-step integrating technology, transducted the H and L chain gene of humanized Fab fragment of anti-HBsAg antibody into the genome of methylotropic yeast P. pastoris. Constructed a engineering yeast to produce humanized Fab fragment of the anti-HBsAg antibody. The Fab fragment was efficiently secreted into the medium at a concentration of 50～80mg/L. The Fab fragment was purified from culturing supernatant of the recombinant yeas by affinity chromatography. The ELISA analysis showed the high affinity of the expressed humanized Fab fragment to the HBsAg.