中国科学C辑(英文版)
中國科學C輯(英文版)
중국과학C집(영문판)
SCIENCE IN CHINA (Series C)
2003年
1期
1-9
,共9页
刘惠%卢芳%陈建军%任红玉%陈常庆
劉惠%盧芳%陳建軍%任紅玉%陳常慶
류혜%로방%진건군%임홍옥%진상경
human tumor necrosis factor%site-directed mutation%cytotoxicity%LD50%Western blot
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.
