Novel affinity tag system using structurally defined antibody-tag interaction: Application to single-step protein purification
Novel affinity tag system using structurally defined antibody-tag interaction: Application to single-step protein purification
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DOI：
10.1110/ps.038299.108
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发表时间：
2008-12-01
期刊：
影响因子：
8
通讯作者：
Takagi, Junichi
中科院分区：
文献类型：
--
作者：
Nogi, Terukazu;Sangawa, Takeshi;Takagi, Junichi
生物学上重要的人类蛋白质通常需要哺乳动物细胞表达用于结构研究，在生产/纯化过程中存在技术和经济问题。我们介绍了一种新的亲和肽标记系统，使用低亲和力的抗肽单克隆抗体。短识别序列的串联使得能够成功地工程化具有理想溶液结合动力学的18个残基的亲和标签，当与通过水混溶性有机溶剂的非变性洗脱相结合时，提供低成本的纯化手段。三维信息为抗体-肽相互作用提供了坚实的结构基础，为进一步改进/修饰提供了机会。
Biologically important human proteins often require mammalian cell expression for structural studies, presenting technical and economical problems in the production/purification processes. We introduce a novel affinity peptide tagging system that uses a low affinity anti-peptide monoclonal antibody. Concatenation of the short recognition sequence enabled the successful engineering of an 18-residue affinity tag with ideal solution binding kinetics, providing a low-cost purification means when combined with nondenaturing elution by water-miscible organic solvents. Three-dimensional information provides a firm structural basis for the antibody-peptide interaction, opening opportunities for further improvements/modifications.