用于重组蛋白质纯化的新材料
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- 英文论文题名：NOVEL MATERIALS FOR HIGHLY SELECTIVE PURIFICATION OF THE RECOMBINANT PROTEIN
- 论文作者：李森武 ; 杨开广 ; 刘路宽 ; 张丽华 ; 张玉奎
- 英文论文作者：Senwu Li ; Kaiguang Yang ; Lukuan Liu ; Lihua Zhang ; Yukui Zhang ; National Chromatographic R.& A.Center ; Key Laboratory of Separation Science for Analytical Chemistry ; Dalian Institute of Chemical Physics ; Chinese Academy of Sciences
- 年：2016
- 作者机构：中国科学院大连化学物理研究所中科院分离分析化学重点实验室国家色谱研究分析中心;
- 论文关键词：重组蛋白质 ; 纯化 ; IMAC ; 分子印迹 ; 活性可控自由基聚合
- 会议召开时间：2016-07-01
- 会议录名称：中国化学会第30届学术年会摘要集-第二十三分会：复杂样品分离分析
- 语种：中文
- 分类号：O629.73
- 学会代码：ZGHY
- 会议名称：中国化学会第30届学术年会-第二十三分会 ; 复杂样品分离分析
- 会议地点：中国辽宁大连
- 主办单位：中国化学会
- 学会名称：中国化学会
- 页数：2
- 文件大小：194k
- 原文格式：D
- 会议级别：全国
摘要
重组蛋白技术在蛋白质结晶、蛋白质药物、蛋白质相互作用及结构蛋白质组学研究中具有非常重要的作用。当前对重组蛋白的纯化,目前常采用固定化金属离子亲和色谱(IMAC)技术,即通过IMAC材料上的金属离子与组氨酸标签之间的螯合作用实现带组氨酸标签的重组蛋白的纯化,但是由于IMAC材料上金属离子暴露在材料表面,任何能与金属离子产生螯合作用的蛋白质均被捕获在IMAC上,如表面富含组氨酸、半胱氨酸、赖氨酸的蛋白质、以及含有金属离子的蛋白质。这些蛋白质的吸附会导致重组蛋白质纯度大幅降低。为了解决这一问题,我们制备了两类新材料,用于提高重组蛋白质纯度:1)通过分子印迹技术,以组氨酸标签为模板,在IMAC材料表面形成组氨酸标签的分子印迹层,使得不含组氨酸标签的蛋白质无法靠近IMAC材料,从而提高重组蛋白纯度;2)通过活性可控自由基聚合,在IMAC基质材料表面包被了一层尺寸可控的聚合物涂层。在聚合物网络的筛分作用下,进而实现带组氨酸标签的重组蛋白的纯化。
Recombinant protein technology occupies an important position in the fields including biopharmaceutics,proteomics,structural and functional biology.However,the purification of recombinant protein is dragged seriously by the impurities.To remove the barrier,two novel purification materials were prepared to enhance the selectivity of IMAC.In the study of epitope surface imprinted materials,the novel epitope imprinting enhanced IMAC(EI-IMAC) showed an obviously improvement on the selectivity of His-tagged recombinant protein from the crude cell lysis.In the study of synergistic sieving immobilized metal-ion affinity chromatograph(SIMAC),a novel tailor-made seperation material for the His-tagged proteins was proposed,which employed the synergistic effect of size sieving effect and metal chelating affinity.Benefitting from the exclusion of interference host proteins by the polymer-based sieving effect,SIMAC could purify the His-tagged protein with high selectivity.
Recombinant protein technology occupies an important position in the fields including biopharmaceutics,proteomics,structural and functional biology.However,the purification of recombinant protein is dragged seriously by the impurities.To remove the barrier,two novel purification materials were prepared to enhance the selectivity of IMAC.In the study of epitope surface imprinted materials,the novel epitope imprinting enhanced IMAC(EI-IMAC) showed an obviously improvement on the selectivity of His-tagged recombinant protein from the crude cell lysis.In the study of synergistic sieving immobilized metal-ion affinity chromatograph(SIMAC),a novel tailor-made seperation material for the His-tagged proteins was proposed,which employed the synergistic effect of size sieving effect and metal chelating affinity.Benefitting from the exclusion of interference host proteins by the polymer-based sieving effect,SIMAC could purify the His-tagged protein with high selectivity.
引文
[1]Li,S.;Yang,K.;Zhao,B.;Li,X.;Liu,L.;Chen,Y.;Zhang,L.;Zhang,Y.;J.Mater.Chem.B 2016,4:1960.
[2]Li,S.;Yang,K.;Deng,N.;Min,Y.;Liu,L.;Zhang,L.;Zhang,Y.;ACS Appl.Mater.Interfaces 2016,8:5747.
[3]Li,S.;Yang,K.;Liu,J.;Jiang,B.;Zhang,L.;Zhang,Y.;Anal.Chem.2015,87:4617.
[2]Li,S.;Yang,K.;Deng,N.;Min,Y.;Liu,L.;Zhang,L.;Zhang,Y.;ACS Appl.Mater.Interfaces 2016,8:5747.
[3]Li,S.;Yang,K.;Liu,J.;Jiang,B.;Zhang,L.;Zhang,Y.;Anal.Chem.2015,87:4617.