Chem. J. Chinese Universities ›› 2009, Vol. 30 ›› Issue (8): 1586.

• Articles • Previous Articles     Next Articles

Primary Studies on the Properties of Trichosanthin After Site-directed Mutagenesis and PEGylation

AN Qun-Xing1,2, LEI Ying-Feng2, YANG Jing2, LI Zhi-Dong2, MU Shi-Jie1*, XU Zhi-Kai2*   

  1. 1. Department of Blood Transfusion, Xijing Hospital, Fourth Military Medical University, Xi′an 710032, China;
    2. Department of Microbiology, Fourth Military Medical University, Xi′an 710033, China
  • Received:2009-03-11 Online:2009-08-10 Published:2009-08-10
  • Contact: XU Zhi-Kai. E-mail: zhikaixu@fmmu.edu.cn; MU Shi-Jie. E-mail: bestar01@163.com
  • Supported by:

    国家自然科学基金(批准号: 30571675)和西京医院学科助推计划(批准号: XJZT07M09)资助.

Abstract:

To construct a PEGylated trichosanthin(TCS) mutein by site-directed PEGylation and analyse its bioactivities, immunogenicity, acute toxicity and pharmacokinetics. By computer modeling, the site KR173-174 was identified as potential antigenic determinant of TCS. Then, a TCS mutein namely TCSKR173-174CG was constructed by site-directed mutagenesis and expressed in E.coli BL21(DE3). After that, the bioactivities, immunogenicity, acute toxicity and pharmacokinetics of the mutant TCS and PEGylated TCS were examined. It was shown that the bioactivities of the mutant TCS were similar to wild-type TCS(wTCS), but with much lower immunogenicity. The PEGylated TCS show a significant decrease in immunogenicity and acute toxicity, and a marked increase in plasma half-life when compared with unmodified TCS. But there is a problem of activity reduction. The site-directed mutagenesis and PEGylation of TCS provide a new approach for reconstructing TCS.

Key words: Trichosanthin; Site-directed PEGylation; Immunogenicity; Pharmacokinetics; Human immunodeficiency virus

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