抗RBC/HBsAg ds-diabody包含体复性条件的优化  

Optimization of Condition for Refolding of Anti-RBC/HBsAg ds-Diabody Inclusion Body

作　　者：陈宇萍[1] 乔媛媛[1] 解鹏[1] 何立东[1] 王琰[1] 

机构地区：[1]海军总医院基础医学研究中心,北京100048

出　　处：《中国生物制品学杂志》2010年第9期1005-1008,共4页Chinese Journal of Biologicals

基　　金：全军医药卫生重点项目(96Z004);博士后科学基金(2004035227)

摘　　要：目的优化以包含体形式表达的抗RBC/HBsAg ds-diabody的复性条件。方法在大肠杆菌BL21/λDE3中诱导表达抗RBC/HBsAg ds-diabody,收获包含体,变性后,分别经透析复性和稀释复性方法复性,并以两种特异性结合活性检测复性效果,对pH值、复性时间和温度进行优化。结果包含体经变性后,pTH-dsR30KD特异蛋白占总蛋白的85%,pTdsR-H30KD特异蛋白占总蛋白的96%;ds-diabody抗体以稀释复性效果较好;在碱性(pH8.8～10)、低温(4℃)条件下复性72h效果最好。结论优化了抗RBC/HBsAg ds-diabody包含体的复性条件。Objective To optimize the condition for refolding of anti-RBC/HBsAg ds-diabody expressed in a form of inclusion body.Methods Anti-RBC/HBsAg ds-diabody was expressed in a form of inclusion body in E.coli BL21/λDE3 under induction of IPTG and folded,then refolded by dialysis and dilution respectively,and the refolding effect was analyzed by determinations of binding activities to HBsAg and RBC,based on which the pH value as wall as time and temperature for refolding were optimized.Results After folding of inclusion body,pTH-dsR 30 KD and pTdsR-H 30 KD specific proteins accounted for 85% and 96% of total protein respectively.The refolding effect of ds-diabody by dilution was satisfactory.The optimal pH value,temperature and time for refolding were 8.8 ~ 10,4℃ and 72 h respectively.Conlusion The condition for refolding of anti-RBC/HBsAg ds-diabody was optimized.

关 键 词：ds-diabody 包含体 蛋白质复性 

分 类 号：Q819[生物学—生物工程] R392.11[医药卫生—免疫学]