BTag: A novel six-residue epitope tag for surveillance and purification of recombinant proteins
BTag: A novel six-residue epitope tag for surveillance and purification of recombinant proteins
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DOI：
10.1016/0378-1119(95)00795-4
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发表时间：
1996-02-22
期刊：
影响因子：
3.5
通讯作者：
Eaton, BT
中科院分区：
文献类型：
--
作者：
Wang, LF;Yu, M;Eaton, BT
表位标签技术(Eta)正成为分子生物学和生物技术中检测、鉴定和纯化重组蛋白(Re-Proteins)的一种越来越有用的技术。在这里，我们描述了一个新的Eta系统，它由两个不同的单抗(mAb；D11和F10)和一个6-氨基酸Eta(Gln-Tyr-Pro-Ala-Leu-Thr或QYPALT)组成。该Eta来源于蓝舌病病毒主要核心蛋白VP7的一个高度保守的区域，因此被命名为BTag。BTag在目前的标签系统中是独一无二的，因为它缺乏电荷，而且标签序列可以放置和检测到再蛋白的任何区域。BTag的其他有用特征包括它的小尺寸和它可以被两种不同的mAb识别。使用BTag系统，已经从各种宿主生物中产生了30多个Re蛋白，并且到目前为止测试的所有蛋白中都保持了标签序列的抗原性。我们的结果表明，在某些应用中，BTag可能优于其他现有的Eta系统。
Epitope tagging (Eta) is becoming an increasingly useful technique in molecular biology and biotechnology for the detection, characterisation and purification of recombinant proteins (re-proteins). Here we describe a novel Eta system composed of two different monoclonal antibodies (mAb; D11 and F10) and a 6-amino-acid Eta (Gln-Tyr-Pro-Ala-Leu-Thr or QYPALT). This Eta was derived from a highly conserved region of the major core protein, VP7, of bluetongue (BT) viruses, hence the name BTag. BTag is unique among current tagging systems in its lack of charge and the fact the tag sequence can be placed and detected in any region of a re-protein. Other useful features of BTag include its small size and its recognition by two different mAb. Using the BTag system, more than 30 re-proteins have been produced from a variety of host organisms, and the antigenicity of the tag sequence was maintained in all of the proteins tested to date. Our results demonstrated that BTag could be superior to other existing Eta systems for certain applications.