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Int J Biol Sci 2011; 7(5):618-628. doi:10.7150/ijbs.7.618

Research Paper

Expression, Purification and Characterization of A Novel Soluble Human Thymosin Alpha1 Concatemer Exhibited A Stronger Stimulation on Mice Lymphocytes Proliferation and Higher Anti-tumor Activity

Weina Li1,*, Liqiang Song3,*, Shouzhen Wu1, Xiaochang Xue1, Lu Zhang2, Liqing He1, Wei Han1, Qing Wang4, Rui Ling 1, Wei Zhang 1, Zhen Yan2 ✉, Yingqi Zhang1 ✉

1. The State Key Laboratory of Cancer Biology, Department of Biopharmaceutics, School of Pharmacy, Fourth Military Medical University, 710032 Xi'an, China
2. The State Key Laboratory of Cancer Biology, Department of Pharmacogenomics, School of Pharmacy, Fourth Military Medical University, 710032 Xi'an, China
3. Department of Respiratory Medicine, Xijing Hospital, Fourth Military Medical University, 710032 Xi'an, China
4. Department of General Surgery, Tangdu Hospital, Fourth Military Medical University, 710038 Xi'an, China
* These authors contributed equally to this article.

Abstract

Thymosin alpha 1 (Tα1) has immunomodulatory and anti-tumor effects in patients and has been commercialized in worldwide. An innovative technique is therefore impending to achieve high-yield expression and purification of Tα1 to meet the increasing requirements for clinical applications. Tα1 can enhance T cells, dendritic cells and antibody responses, and also augment an anti-tumor immune response. In the current study, we developed a novel technique to produce Tα1 concatemer and investigated its capability in anti-tumor immunotherapy. We expressed the recombinant 2×Tα1 concatemer protein (Tα1② protein) in Escherichia coli. The purity of Tα1② was higher than 95% as assessed by HPLC analysis. In vitro, Tα1② could stimulate the proliferation of mouse splenic lymphocyte, and increase the apoptosis of tumor cell lines. In vivo, Tα1② significantly inhibited the tumor growth in B16 tumor-bearing mice. Compared with Tα1, the Tα1② is of more effective bioactivity than Tα1. The purified Tα1② is a promising substitute for synthetic Tα1 because of its potent anti-tumor effects. We concluded that the expression system for Tα1 concatemer was constructed successfully, which could serves as a highly efficient tool for the production of large quantities of the highly active protein.

Keywords: Thymosin alpha 1, concatemer, protein expression, purification, tumor suppression

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How to cite this article:
Li W, Song L, Wu S, Xue X, Zhang L, He L, Han W, Wang Q, Ling R, Zhang W, Yan Z, Zhang Y. Expression, Purification and Characterization of A Novel Soluble Human Thymosin Alpha1 Concatemer Exhibited A Stronger Stimulation on Mice Lymphocytes Proliferation and Higher Anti-tumor Activity. Int J Biol Sci 2011; 7(5):618-628. doi:10.7150/ijbs.7.618. Available from http://www.ijbs.com/v07p0618.htm