亲和分离材料的设计策略及其应用
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- 英文论文题名：Design and Application of Affinity Separation Materials
- 论文作者：贾凌云 ; 任军 ; 臧柏林 ; 暴晓博
- 英文论文作者：Lingyun Jia ; J Ren ; B Zang ; B Bao ; School of life science and biotechnology ; Dalian University of Technology
- 年：2016
- 作者机构：大连理工大学生命科学与技术学院;
- 论文关键词：亲和色谱 ; 纳米抗体 ; 定向固载 ; 超分子洗脱
- 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：TB383.1;O658
- 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：1
- 文件大小：186k
- 原文格式：D
- 会议级别：全国
摘要
亲和分离材料在蛋白质组学及抗体类药物分离等领域有着广泛的应用,但如何降低亲和分离材料在复杂样品分离分析中的非特异吸附、提高检测的灵敏度、改善纯度和活性仍然是材料制备过程中关注的核心问题。近年来,针对复杂血液样品中低丰度蛋白的吸附,我们从基因工程菌的构建出发,建立了目标蛋白单链纳米抗体定向固载策略,在体外实现了甲酰甘氨酸生成酶对纳米抗体碳端修饰酶识别肽序列的有效转化,制备了纳米抗体定向固载亲和分离材料,实现了对血液中微克级β2微球蛋白95%以上的吸附选择性。针对高丰度抗体,通过对材料空间结构和活化密度的有效控制,优化了疏水电荷诱导层析亲和色谱材料制备方法,对血液中抗体的吸附选择性达到85%以上。提出了基于超分子作用的置换色谱模式并应用于抗体分离,在生理条件下,利用15 mM的β环糊精实现了抗体的有效洗脱,摆脱了抗体色谱分离过程中高盐和低pH值的苛刻洗脱条件,有效保持了抗体活性。
Affinity separation materials have been extensively used in proteomics,purification of antibody drugs and so on.However,the problem such as how to decrease the non-specific adsorption,promote the sensitivity and increase the purity and activity of product are still the key point in the fabrication of those materials.Recently,to adsorb the low-abundance protein in blood,we developed a strategy for site-specific immobilization of single-domain antibodies(sdAb).Additionally,we prepared affinity separation materials with site-specifically immobilized sdAb,achieving 95%of adsorption specificity to nanomolar β2-microglobulin in blood.To remove the high abundance protein such as antibody,we optimized the preparation of the matrix for hydrophobic charge induction chromatography,achieving 85%of adsorption specificity to IgG in blood.Meanwhile,we established the salt-independent hydrophobic displacement chromatography for antibody purification using cyclodextrin as supermolecular displacer,which are independent of high concentrations of salt or pH alternation in elution step.
Affinity separation materials have been extensively used in proteomics,purification of antibody drugs and so on.However,the problem such as how to decrease the non-specific adsorption,promote the sensitivity and increase the purity and activity of product are still the key point in the fabrication of those materials.Recently,to adsorb the low-abundance protein in blood,we developed a strategy for site-specific immobilization of single-domain antibodies(sdAb).Additionally,we prepared affinity separation materials with site-specifically immobilized sdAb,achieving 95%of adsorption specificity to nanomolar β2-microglobulin in blood.To remove the high abundance protein such as antibody,we optimized the preparation of the matrix for hydrophobic charge induction chromatography,achieving 85%of adsorption specificity to IgG in blood.Meanwhile,we established the salt-independent hydrophobic displacement chromatography for antibody purification using cyclodextrin as supermolecular displacer,which are independent of high concentrations of salt or pH alternation in elution step.
引文
[1]Zang,B.;Ren,J;Xu,L;Jia,L*.J Chromatogr B,2016,1008:132.
[2]Ren,J.;Yao,P.;Chen,J.;Jia,L*.J Chromatogr A,2014,1369:98.
[3]Xu,L.;Wang,C.;Chen,L.;Ren,J.;Xie,J.;Jia,L*.J Chromatogr B,2014,971:94
[4]Ren,J.;Yao,P.;Cao,Y.;Cao,J.;Zhang,L.;Wang,Y.;Jia,L*.J Chromatogr A,2014,1352:62
[2]Ren,J.;Yao,P.;Chen,J.;Jia,L*.J Chromatogr A,2014,1369:98.
[3]Xu,L.;Wang,C.;Chen,L.;Ren,J.;Xie,J.;Jia,L*.J Chromatogr B,2014,971:94
[4]Ren,J.;Yao,P.;Cao,Y.;Cao,J.;Zhang,L.;Wang,Y.;Jia,L*.J Chromatogr A,2014,1352:62