突变型(α99,β82位Lys-Cys)人重组血红蛋白的纯化  

Purification of α99,β82 Recombinant Human Mutant (α99,β82 Lys-Cys)Hemoglobin

作　　者：张浩[1] 毛秉智[1] 王全立[2] 冯起 刘晓达[2] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850 [2]军事医学科学院输血研究所,北京100850

出　　处：《药物生物技术》2000年第4期200-203,共4页Pharmaceutical Biotechnology

摘　　要：将基因突变型 (α99,β82位Lys Cys)血红蛋白在 pBV2 2 0载体中诱导表达 ,表达产物达细菌总蛋白的 2 0 %左右。该表达产物以包涵体形式存在 ,包涵体经洗涤后 ,用 8mol/L尿素溶解 ,先用Q SepharoseFastFlow阴离子交换纯化 ,再经Sephacryl 10 0凝胶过滤纯化。纯化产物经复性 。Recombinant human α99,β82 mutant genes were introduced into the expression vector pBV220 The expression product reached about 20% of total bacterial protein They were expressed as inclusion bodies and must be dissolved in denatured condition and then renatured After the washing by 4 mol/L urea,purification was performed on a column of Q sepharose Fast Flow Further purification was performed on a column of Sephacryl 100 gel filtration which resulted in the elimination of most of the contaminated proteins Then a refolding reaction was established in the presence of glutathione oxide shuffling system The purified and renatured product had biological activity

关 键 词：纯化 人重组血红蛋白 基因重组 血液制剂 

分 类 号：TQ464.56[化学工程—制药化工]