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镍离子金属螯合亲和层析纯化重组类人胶原蛋白

发表时间:2013-03-15  浏览量:1030  下载量:320
全部作者: 乔媛媛,范代娣,邓建军,惠俊峰,朱晨辉,马晓轩
作者单位: 西北大学化工学院;西北工业大学,陕西省可降解生物医用材料重点实验室
摘 要: 以Ni2+快流速琼脂糖凝胶为层析介质,分别采用咪唑洗脱和降低pH值洗脱方式对重组类人胶原蛋白(recombinant human-like collagen,RHLC)进行纯化,收集洗脱部分,SDS-PAGE检测蛋白纯度。结果表明:以含20 mmol/L NaH2PO4, 0.5 mol/L NaCl(pH= 4.0)的洗脱缓冲液进行降低pH值洗脱所得蛋白纯度和收率最好,所得蛋白纯度达99%以上,回收率为76%. 纯化的RHLC为电泳纯,分子量为97 ku.
关 键 词: 蛋白质工程;金属螯合亲和层析;类人胶原蛋白;纯化
Title: Purification of recombinant human-like collagen by Ni(II)-based immobilized metal ion affinity chromatography
Author: QIAO Yuanyuan, FAN Daidi, DENG Jianjun, HUI Junfeng, ZHU Chenhui, MA Xiaoxuan
Organization: College of Chemical Engineering, Northwest University; Shaanxi Key Laboratory of Degradable Biomedical Materials, Northwest Unirersity
Abstract: In this paper, the recombinant human-like collagen (RHLC) was purified by elution methods of iminazole and lower pH respectively with Ni2+ sepharose fast flow as the affinity medium. Various fractions of elution were collected and then protein purity was identified for by SDS-PAGE. The results showed that the purity and the recovery of the RHLC is the best by the lower pH elution with the buffer (20 mmol/L NaH2PO4, 0.5 mol/L NaCl, pH=4.0). The protein purity was more than 99%, and the recovery rate was 76%. The purified RHLC was electrophoretically pure with a relative molecular weight of 97 ku.
Key words: protein engineering; immobilized metal ion affinity chromatography; recombinant human-like collagen; purification
发表期数: 2013年3月第5期
引用格式: 乔媛媛,范代娣,邓建军,等. 镍离子金属螯合亲和层析纯化重组类人胶原蛋白[J]. 中国科技论文在线精品论文,2013,6(5):402-406.
 
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