| Construction of novel tumor necrosis factor-alpha mutants with reduced toxicity and higher cytotoxicity on human tumor cells |
| |
| 作者姓名： | 刘惠 卢芳 陈建军 任红玉 陈常庆 |
| |
| 作者单位： | Shanghai Research Center of Biotechnology，Chinese Academy of Sciences，Shanghai Research Center of Biotechnology，Chinese Academy of Sciences，Shanghai Research Center of Biotechnology，Chinese Academy of Sciences，Shanghai Research Center of Biotechnology，Chinese Academy of Sciences，Shanghai Research Center of Biotechnology，Chinese Academy of Sciences Shanghai 200233，China，Shanghai 200233，China，Shanghai 200233，China，Shanghai 200233，China，Shanghai 200233，China |
| |
| 摘 要： | Theremarkableabilityofhumantumornecrosisfactor(hTNF-a)istokillmanymalignantcelllinesinvitroorinvivoselectivelyandhavealmostnotoxicityfornormaltissuecells1,2].However,manysideeffectsofhTNF-ainclinictrialshaveseverelylimiteditsapplicationincancertreatment3].Recently,alotofworkhasbeendoneforimprovinghTNF-abymeansofproteinengineeringtoobtainnovelhTNF-amutantswithhighcytotoxcityandreducedsystematictoxicity.Yamagishietal.pointedoutthattheessentialfourregionsformaintainingtheactivityofhTNF-aw…
|
| 收稿时间： | 2001-05-29 |
Construction of novel tumor necrosis factor-alpha mutants with reduced toxicity and higher cytotoxicity on human tumor cells |
| |
| LIU Hui,LU Fang,CHEN Jianjun,REN Hongyu & CHEN Changqing Shanghai Research Center of Biotechnology,Chinese Academy of Sciences,Shanghai ,China Correspondence should be addressed to Chen Changqing.Construction of novel tumor necrosis factor-alpha mutants with reduced toxicity and higher cytotoxicity on human tumor cells[J].Science China Life sciences,2003,46(1):1-9. |
| |
| Authors: | LIU Hui LU Fang CHEN Jianjun REN Hongyu & CHEN Changqing Shanghai Research Center of Biotechnology Chinese Academy of Sciences Shanghai China Correspondence should be addressed to Chen Changqing |
| |
| Institution: | email: cqchen@server.shcnc.ac.cn |
| |
| Abstract: | Two tumor necrosis factor-( mutants MT1 (32Trp157Phe) and MT2 (2Lys30Ser- 32Trp157Phe) were constructed by site-directed mutagenesis. These mutants were soluble and over-expressed in E. coli. The purity of purified mutants was above 95% by serial chromatography. The results of Western blot indicated that these mutants could be cross-reactive with monoclonal antibody against native hTNF-α. Compared to parent hTNF-α, the cytotoxicity of these mutants on murine fibrosarcoma L929 cell lines reduced 4-5 orders of magnitude but was equivalent to that of native hTNF-α on human tumor cell lines. The LD50 of mutant MT1 was reduced to 0.34% of wild type and the dose of MT2 that resulted in 30% death of mice reduced to less than 1/700 that of parent hTNF-α. |
| |
| Keywords: | human tumor necrosis factor site-directed mutation cytotoxicity LD50 Western blot |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
| 点击此处可从《中国科学：生命科学英文版》浏览原始摘要信息 |
| 点击此处可从《中国科学：生命科学英文版》下载免费的PDF全文 |
