Institute of Molecular & Experimental Therapeutics, East China Normal University, Shanghai, China.
J Chromatogr A. 2011 May 6;1218(18):2553-60. doi: 10.1016/j.chroma.2011.02.053. Epub 2011 Mar 10.
Purification tags are robust tools that can be used to purify a variety of target proteins. However, tag removal remains an expensive and significant issue that must be resolved. Based on the affinity and the trans-splicing activity between the two domains of Ssp DnaB split-intein, a novel approach for tag affinity purification of recombinant proteins with controllable tag removal by inducible auto-cleavage has been developed. This system provides a new affinity method and avoids premature splicing of the intein fused proteins expressed in host cells. The affinity matrix can be reused. In addition, this method is compatible with his-tag affinity purification technique. Our methods provide the insights for establishing a novel recombinant protein preparation system.
纯化标签是强大的工具，可用于纯化各种目标蛋白。然而，标签去除仍然是一个昂贵且重要的问题，必须加以解决。基于 Ssp DnaB 分裂内含子两个结构域之间的亲和性和转剪接活性，开发了一种通过诱导自动切割可控去除标签的重组蛋白标签亲和纯化的新方法。该系统提供了一种新的亲和方法，避免了在宿主细胞中表达的内含子融合蛋白的过早剪接。亲和基质可以重复使用。此外，该方法与 his 标签亲和纯化技术兼容。我们的方法为建立新型重组蛋白制备系统提供了思路。
