重组人白介素6受体功能区片段的功能鉴定

Functional Identification for Recombinant Functional Fragment of Human Interleukin 6 Receptor

用生物素标记重组人白介素6受体功能区片段rIL6R-28及其二联体蛋白rIL6R-53．竞争ELISA表明重组蛋白可以与配基IL-6特异结合．流式细胞术检测结果表明IL-6与生物素标记的重组蛋白所形成的复合物能够与7TD1细胞表面的gp130结合．而7TD1细胞生长分析则表明，重组蛋白可以增强IL-6对7TD1．细胞的生长刺激作用．

The recombinant functional fragment of human IL-6R (rIL6R-28) and its dimer (rIL6R-53)were labelled with biotin. To detect the specific binding of recombinant proteins to IL-6, competitive ELISA was established. Flow cytometry was employed to examine whether or not the complex of IL-6 and the biotinylated recombinant proteins could bind to the surface of 7TD1 cells. Judged by the alteration of fluorescence intensity and positive cell ratio, it was confirmed that the complex of IL-6 and recombinant proteins could bind to gp130. Finally,7TD1 cell growth assay showed that the recombinant proteins were capable of augmenting the growth-stimulating effect of IL-6 on 7TD1 cells. These results suggest that the cytokine receptor family domain is responsible for the function of IL-6R, and the Ig-like domain is required neither for IL-6-binding nor for mediating the IL-6 signal. Besides, the glycosylation seems not to be a determinant factor for IL-6R function.