Construction of immune inhibitor of new rhTNF-?
- VernacularTitle:新型肿瘤坏死因子免疫抑制剂的克隆和表达
- Author:
Wei HAN
;
Ning ZHAO
;
Jihong SHI
- Publication Type:Journal Article
- Keywords:
New rhTNF-?;
Chicken egg-white lysozyme;
Sequence of T-help cell;
Immune inhibitor
- From:
Chinese Journal of Immunology
1985;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct an immune inhibitor of new TNF-?,C-terminal sequence of new rhTNF-? was replaced with sequence of T-help cell of hen egg-white lysozyme(HEL).Methods:The rhTNF-? mutant was cloned 、expressed and purified.Results:The DNA sequencing analysis showed that the C-terminal sequence of new rhTNF-? mutant was correct.The mutant was inserted into pBV220 expression vector .After the recombinant bacteria was incubated at 42℃ for 4 h,a new band of the protein with relative molecular weight of 1.7 kD was shown on the gel.The band amounted to 30 % of total bacteria protein.Western blot showed that the mutant protein could associated with anti-TNF-? antibody.After the protein was purified by through a column of Q-Sepharose Fast Flow, the purity of the protein was above 90%.The biological activity of the protein was measured with L929 cells.The result showed that biological activity of the protein was totally lost.Conclusion:The experimental evidence demonstrated that the construction of new rhTNF-? mutant was successful.The mutant not only can be associated with anti-TNF-? antibody, but also lose the biological activity of original TNF-?. [
