Soluble expression of recombinant human CD137 ligand in Escherichia coli by co-expression of chaperones
Soluble expression of recombinant human CD137 ligand in Escherichia coli by co-expression of chaperones
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通过共表达分子伴侣在大肠杆菌中可溶性表达重组人 CD137 配体
DOI：
10.1007/s10295-011-1045-1
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发表时间：
2012-03
影响因子：
3.4
通讯作者：
Chen, Yijun
中科院分区：
文献类型：
--
作者：
Wang, Shuzhen;Tan, Aimin;Lv, Junfang;Wang, Peng;Yin, Xiaojin;Chen, Yijun
CD 137配体（CD 137 L）是肿瘤坏死因子超家族的成员，其结合CD 137以提供用于T细胞活化的阳性共刺激信号。通过CD 137/CD 137 L共刺激已成为肿瘤治疗的有前途的方法之一。以往的研究表明，在大肠杆菌中表达CD 137 L会导致包涵体或产量低。本研究探讨并分析了5种不同分子伴侣对重组人CD 137 L蛋白可溶性表达的影响。CD 137 L在E.通过伴侣蛋白GroES-GroEL-Tf的共表达，显著提高了大肠杆菌的表达量。经双重诱导和亲和层析后，以3 mg蛋白/L的产量获得纯化的重组CD 137 L，其纯度从原始检测不到的水平大于98%。纯化的重组CD 137 L能与CD 137阳性细胞结合，并呈剂量依赖性，能显著促进活化小鼠T细胞的生长和IL-2的释放。本研究为人共刺激分子CD 137 L的可溶性表达提供了一个有效的系统，为重组蛋白药物的临床开发提供了有利条件。
CD137 ligand (CD137L) is a member of the tumor-necrosis factor superfamily that binds CD137 to provide positive co-stimulatory signals for T cells activation. Co-stimulation through CD137/CD137L has become one of the promising approaches for cancer therapy. Previous reports have shown that CD137L expressed in Escherichia coli resulted in inclusion bodies or low yield. In this study, the effects of five different chaperone teams on the soluble expression of recombinant human CD137L protein were explored and analyzed. The poor expression of CD137L in the cytoplasm of E. coli was improved significantly by co-expression of chaperone GroES-GroEL-Tf. After dual induction and affinity chromatography, purified recombinant CD137L was obtained at a yield of 3 mg protein per liter with purity greater than 98% from original undetectable level. Additionally, the purified recombinant CD137L could bind CD137-positive cells in a dose-dependent manner, markedly promote the growth of activated mice T cells, and elevate the release of IL-2. The present work provides an effective system for soluble expression of functional human co-stimulatory molecule CD137L, which will facilitate the clinical developments of recombinant protein drugs.
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基于伴侣的程序，以增加大肠杆菌中产生的可溶性重组蛋白的产量。
影响因子：
3.5
作者：
通讯作者：
--
影响因子：
2.2
作者：
Erdile, LF;Smith, D;Berd, D
通讯作者：
Berd, D
影响因子：
4.4
作者：
Kazuyo Nishihara;M. Kanemori;M. Kitagawa;H. Yanagi;T. Yura
通讯作者：
Kazuyo Nishihara;M. Kanemori;M. Kitagawa;H. Yanagi;T. Yura
影响因子：
5.5
作者：
Schabowsky RH;Elpek KG;Madireddi S;Sharma RK;Yolcu ES;Bandura-Morgan L;Miller R;MacLeod KJ;Mittler RS;Shirwan H
通讯作者：
Shirwan H
影响因子：
11.2
作者：
Sharma RK;Schabowsky RH;Srivastava AK;Elpek KG;Madireddi S;Zhao H;Zhong Z;Miller RW;Macleod KJ;Yolcu ES;Shirwan H
通讯作者：
Shirwan H