新型电泳介质和技术的发展与蛋白质组学研究
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- 英文论文题名：Novel electrophoresis media and technique for proteomics research
- 论文作者：肖华 ; 刘臻 ; 孙妍 ; 尚志 ; 樊柳荫 ; 曹成喜
- 英文论文作者：Hua Xiao ; Zhen Liu ; Yan Sun ; Zhi Shang ; Liu-Yin Fan ; Cheng-Xi Cao ; School of Life Sciences and Biotechnology ; Shanghai Jiao Tong University
- 年：2016
- 作者机构：上海交通大学生命科学技术学院;
- 论文关键词：凝胶电泳 ; 等电聚焦电泳 ; 蛋白质组学
- 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：O658.9;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：1
- 文件大小：171k
- 原文格式：D
- 会议级别：全国
摘要
凝胶电泳是蛋白质与核酸等大分子分离的重要工具,广泛应用于生物医药领域。虽然凝胶制备技术已经非常成熟,但在复杂蛋白样品的分离方面仍然面临许多挑战,往往存在分辨率低、柔韧性差等缺点。针对传统聚丙烯酰胺凝胶,我们将丙烯酸既作为单体又作为交联剂,合成了新型多共价聚丙烯酰胺凝胶,根据凝胶孔结构的变化提出多共价交联理论,最终在不提高丙烯酰胺比例的情况下显著提高了复杂蛋白样品的分离分辨率,同时增强了凝胶的柔韧性。常规等电聚焦电泳存在pH梯度难以调节的问题,通过相关机理研究,我们基于移动反应界面理论发展了pH梯度可调的新技术,用于双向凝胶电泳显著提高了蛋白质分离分析的分辨率和灵敏度。将新型电泳介质和可调谐的等电聚焦电泳用于蛋白质组学研究,有助于评价样品前处理效果,有利于发现新的蛋白质标志物,显著提高了凝胶电泳的灵活性和实用性。
Gel electrophoresis is a powerful tool for the separation of proteins and nucleic acids.It has been widely used in the field of biomedicine while suffered from limited resolution and flexibility.We in-situ copolymerized a multiple covalent crosslink gel of poly(acrylamide-acrylic acid)(P(AM-AA)),in that acrylic acid and acrylamide were used both as a monomer and as a covalent cross-linker.The resultant gel demonstrated remarkable separation capacity of macromolecular proteins.We also developed novel electrophoresis technique to adjust the pH gradient distribution of conventional isoelectric focusing through the moving reaction boundary theory.Based on the new strategy,the resolution,sensitivity and utility of two-dimensional gel electrophoresis were improved dramatically.The application of these electrophoretic media and technique for proteomics will benefit sample quality evaluation as well as the discovery of novel biomarkers.
Gel electrophoresis is a powerful tool for the separation of proteins and nucleic acids.It has been widely used in the field of biomedicine while suffered from limited resolution and flexibility.We in-situ copolymerized a multiple covalent crosslink gel of poly(acrylamide-acrylic acid)(P(AM-AA)),in that acrylic acid and acrylamide were used both as a monomer and as a covalent cross-linker.The resultant gel demonstrated remarkable separation capacity of macromolecular proteins.We also developed novel electrophoresis technique to adjust the pH gradient distribution of conventional isoelectric focusing through the moving reaction boundary theory.Based on the new strategy,the resolution,sensitivity and utility of two-dimensional gel electrophoresis were improved dramatically.The application of these electrophoretic media and technique for proteomics will benefit sample quality evaluation as well as the discovery of novel biomarkers.
引文
[1]Liu,Z.,Fan,L.Y.,Xiao,H.,Cao,C.X.,Chem.Commun.2016,52:3247-3250
[2]Guo,C.G.,Shang,Z.,Yan,J.,Li,S.,et al.Talanta 2015,137:197-203
[3]Xiao,H.,Zhang,Y.,Kim,Y.,Kim,S.,et al.Sci.Rep.2016,6:22165
[4]Sun,Y.,Xia,Z.J.,Shang,Z.,Sun,K.B.,et al.Sci.Rep.2016,6:24669.
[2]Guo,C.G.,Shang,Z.,Yan,J.,Li,S.,et al.Talanta 2015,137:197-203
[3]Xiao,H.,Zhang,Y.,Kim,Y.,Kim,S.,et al.Sci.Rep.2016,6:22165
[4]Sun,Y.,Xia,Z.J.,Shang,Z.,Sun,K.B.,et al.Sci.Rep.2016,6:24669.