摘要
目的 建立聚乙二醇化重组人干扰素α2b(PEG rhIFNα2b)的质量检测方法。方法 采用Wish细胞 VSV病毒系统 ,以CPE法测定干扰素的生物学活性 ,基质辅助激光解吸附飞行时间质谱法测定相对分子质量 ,反向高效液相法 (RP HPLC)测定纯度 ,溴化氰裂解 SDS PAGE法测定肽图 ,等电聚焦电泳法测定等电点 ,紫外分光光度法测定紫外吸收光谱。结果 聚乙二醇化重组人干扰素α2b的比活为 6 . 0× 10 6~ 10 . 0× 10 6IU/mg蛋白 ,相对分子质量约为 6 2 6 0 0 ,等电点在 5 . 2 0~ 6. 5 5之间 ,紫外最大吸收峰约在 2 78nm波长处。结论 所建立的检测方法可控制聚乙二醇化重组人干扰素α2b质量。
Objective To develop the methods for quality control of PEG-rhIFNα2b.Methods Determine the biological activity of PEG-rhIFNα2b by CPE method using Wish cell-VSV system,the relative molecular weight by MALDI-TOF-MS,the purity by RP-HPLC,the peptide map by SDS-PAGE after lysis with cyanogens bromide,the isoelectric point by isoelectric focusing electrophoresis and the ultraviolet absorption spectrum by ultraviolet spectroscopy.Results The specific activity,relative molecular weight and isoelectric point of PEG-rhIFNα2b were 6.0×106-10.0×106 IU/mg protein,62 600 and 5.20-6.55 respectively,and its maximum ultraviolet absorption peak appeared at 278 nm.Conclusion The developed methods were suitable for the quality control of PEG-rhIFNα2b.
出处
《中国生物制品学杂志》
CAS
CSCD
2005年第1期48-50,共3页
Chinese Journal of Biologicals