Wang Jing, Yang Zhenhui, Shi Ming, Li Yan, Guo Ning, Shen Beifen, Feng Jiannan
Department of Molecular Immunology, Institute of Basic Medical Sciences, Beijing, 100850, P.R. China.
Biotechnol Lett. 2007 Sep;29(9):1323-7. doi: 10.1007/s10529-007-9402-x. Epub 2007 May 30.
To investigate the function of the N-terminal immunoglobulin (Ig)-like domain of the human interleukin-6 receptor alpha-chain (hIL-6R), we constructed a soluble human interleukin-6 receptor (shIL-6R) (named EC05, amino acids 20-354) and soluble variants of the shIL-6R lacking the Ig-like domain (named EC70, amino acids 105-354). The two extracellular portions of hIL-6R were expressed as soluble fusion proteins with thioredoxin in Escherichia coli and purified by using Ni-NTA agarose. Western blot showed that purified proteins were immunoreactive with the antibody against hIL-6R. They also possessed specific binding activity with human interleukin-6 (hIL-6) in ELISA analysis.
为了研究人白细胞介素-6受体α链（hIL-6R）的N端免疫球蛋白（Ig）样结构域的功能，我们构建了一种可溶性人白细胞介素-6受体（shIL-6R）（命名为EC05，氨基酸20 - 354）以及缺乏Ig样结构域的shIL-6R可溶性变体（命名为EC70，氨基酸105 - 354）。hIL-6R的这两个细胞外部分在大肠杆菌中作为与硫氧还蛋白的可溶性融合蛋白表达，并使用Ni-NTA琼脂糖进行纯化。蛋白质印迹法显示，纯化后的蛋白质与抗hIL-6R抗体发生免疫反应。在酶联免疫吸附测定分析中，它们与人白细胞介素-6（hIL-6）也具有特异性结合活性。
