The Study of Mouse TNF-? Functional Domain and Its Neutralizing Antibody Binding Site
- VernacularTitle:鼠TNF-?功能位点及其中和性抗体结合位点研究
- Author:
Junying JIA
;
Hongzhe ZHOU
;
Jie TANG
- Publication Type:Journal Article
- Keywords:
mouse TNF-?, neutralizing antibody, binding site, yeast display
- From:
Progress in Biochemistry and Biophysics
2006;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Tumor necrosis factor alpha(TNF-?) is a multi-functional cytokine that plays a significant role in many autoimmune diseases. The key biological functional domain of mouse TNF-? was determined by identifying the binding site of mouse TNF-? neutralizing antibody 9C6. Using yeast surface display technology, it was determined that 9C6 can recognize the linear amino acid fragment 29 ～40 of mature mouse TNF-? protein. Through mutagenesis experiments of this TNF-? region, the critical amino acids necessary for 9C6 binding were identified. Finally, wild-type mouse TNF-? and mutant variants that loses binding ability to 9C6 were expressed in Escherichia coli, purified, and used in a L929 cell killing assay. The assay results proved that the key amino acids for 9C6 binding were consistent with mouse TNF-? functional domain.
