抗IV型胶原酶单链抗体在毕赤酵母中分泌表达

EXPRESSION OF SINGLE CHAIN ANTIBODY TO COLLAGENASE IV BY PICHIA PASTORIS

利用毕赤酵母系统表达抗 IV型胶原酶人单链抗体。首先把目的基因克隆到毕赤酵母表达载体上 ,电击转化受体菌。在甲醇诱导下表达单链抗体。 SDS- PAGE和免疫印迹显示毕赤酵母分泌表达人单链抗体 ,表达量约 2 0 mg/ L酵母培养物。该表达系统与大肠杆菌相比 ,简化了表达产物的分离纯化程序。

To express human single chain antibody to collagenase IV as soluble proteins secreted by pichia pastoris, a recombinant vector scFv pPIC9 was constructed, and then transformed into pichia pastoris GS115 by electuoporation, The transformants were selected by DNA hybridization and cultured in the media with methanlo. Soluble scFv were secreted into culturesupernatant and characterized by SDS PAGE and Western Blot. Our results showed that the secreting of soluble scFv in pichia pastoris was as high as 20mg/L. The soluble scFv has similar immuno activity to its counterpart produced in bacteria, and it is more easily and efficiently purified.