重组人白细胞介素6 部分理化生物学性质的鉴定  

Determination of some biochemical and physical properties of rhIL 6

作　　者：陶好霞[1] 边 疆[1] 孙 澎[1] 巩 新[1] 薛 冲[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《军事医学科学院院刊》1999年第3期197-200,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：军事医学科学院开发基金

摘　　要：目的：对 Ｎ 端缺失 ５ 个氨基酸的重组人白细胞介素 ６（ｒｅｃｏｍ ｂｉｎａｎｔ ｈｕｍ ａｎ ｉｎｔｅｒｌｅｕｋｉｎ６，ｒｈ Ｉ Ｌ６）纯品的理化生物学性质进行分析鉴定。方法：用蛋白序列自动分析仪、紫外及可见光扫描仪、 Ｓ Ｄ Ｓ聚丙烯酰胺凝胶电泳、溴化氰裂解、等电聚焦电泳、蛋白质印迹法和 Ｍ Ｔ Ｔ 比色法分别对ｒｈ Ｉ Ｌ６ 缺失体 Ｎ 端氨基酸序列、紫外及可见光吸收光谱、相对分子质量、肽图、等电点（ｐ Ｉ）、抗原特异性及生物学活性进行了分析鉴定。结果：与天然 Ｉ Ｌ６ 相比，ｒｈ Ｉ Ｌ６ Ｎ 端缺失了 ５ 个氨基酸；在２７６ ｎｍ 波长处有一个特异的蛋白吸收峰；相对分子质量为 ２１ ４８７ ；裂解的肽段分布在理论值的肽段分布范围之内；ｐ Ｉ为６．１；能与兔抗人 Ｉ Ｌ６ 抗体特异性结合；比活性达２．０５×１０１１ Ｉ Ｕ／ｇ。结论：缺失 ５ 个氨基酸的 ｒｈ Ｉ Ｌ６ 生物学活性与天然 Ｉ Ｌ６Objective: To characterize the rhIL 6 with 5 amino acids deleted at the N terminal. Methods: The N terminal sequence, ultraviolet and visible spectra, molecular weight, peptide map, isoelectric point (pI), antigen specificity were analyzed with protein sequence autoanalyzing, whole wavelength scanning, SDS PAGE, cyanogen bromide lysis, isoelectric focusing, Western blot and MTT colorimetry respectively. Results: The sequence at the rhIL 6 defective N terminal is MEDSKD...; there is a specific protein absorption peak at 276 nm; its relative molecular weight is 21 487; the lysed peptide segments distributed within that of the range of its theoretical values; its pI is 6.1; it is able to specifically bind with rabbit anti human IL 6 antibody; its specific activity is 2.05×10 11 IU/g. Conclusion: The biological activity of the rhIL 6 deleting 5 amino acids at the N terminal is similar to that of the native IL 6.

关 键 词：白细胞介素6 分子量 等电点 免疫印迹 RHIL-6 

分 类 号：R392.11[医药卫生—免疫学]