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《Progress in Biochemistry and Biophysics》 2009-04
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The Study of Mouse TNF-α Functional Domain and Its Neutralizing Antibody Binding Site

JIA Jun-Ying1,2), ZHOU Hong-Zhe1), TANG Jie1) (1)Institute of Biophysics, The Chinese Academy of Sciences, Beijing 100101, China; 2)Graduate School of The Chinese Academy of Sciences, Beijing 100039, China)  
Tumor necrosis factor alpha(TNF-α) is a multi-functional cytokine that plays a significant role in many autoimmune diseases. The key biological functional domain of mouse TNF-α was determined by identifying the binding site of mouse TNF-α 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-α protein. Through mutagenesis experiments of this TNF-α region, the critical amino acids necessary for 9C6 binding were identified. Finally, wild-type mouse TNF-α 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-α functional domain.
【Fund】: 国家高技术研究发展计划(863)资助项目(2006AA02090805 2006AA02090402)~~
【CateGory Index】: Q51
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