title: 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.
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.
authors.affiliation: The State Key Laboratory of Cancer Biology, Department of Biopharmaceutics, School of Pharmacy, Fourth Military Medical University, 710032 Xi'an, China.
authors.forename: Weina
authors.initials: W
authors.lastname: Li
authors.forename: Liqiang
authors.initials: L
authors.lastname: Song
authors.forename: Shouzhen
authors.initials: S
authors.lastname: Wu
authors.forename: Xiaochang
authors.initials: X
authors.lastname: Xue
authors.forename: Lu
authors.initials: L
authors.lastname: Zhang
authors.forename: Liqing
authors.initials: L
authors.lastname: He
authors.forename: Wei
authors.initials: W
authors.lastname: Han
authors.forename: Qing
authors.initials: Q
authors.lastname: Wang
authors.forename: Rui
authors.initials: R
authors.lastname: Ling
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Zhen
authors.initials: Z
authors.lastname: Yan
authors.forename: Yingqi
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002453:Cell Cycle / Q000187:drug effects*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D018922:HL-60 Cells; D056945:Hep G2 Cells; D006801:Humans; D008213:Lymphocyte Activation / Q000187:drug effects*; D008546:Melanoma, Experimental / Q000188:drug therapy / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D000077596:Thymalfasin; D013947:Thymosin / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology
chemical_list: D013947:Thymosin; D000077596:Thymalfasin