重组人白细胞介素12的纯化与鉴定  

Identification and purification of recombinant human interleukin-12

作　　者：张宝定[1] 程民[1] 肖祥[1] 何海辉[1] 刘文涛[1] 费保珍[1] 田志刚[1] 魏海明[1] 

机构地区：[1]中国科学技术大学生命科学学院免疫学研究所,安徽合肥230027

出　　处：《中国生化药物杂志》2009年第1期14-17,共4页Chinese Journal of Biochemical Pharmaceutics

基　　金：国家863目标导向课题2006AA02Z491资助

摘　　要：目的探索重组人白细胞介素12(rhIL-12)的纯化与鉴定方法。方法采用Millipore超滤浓缩系统对含rhIL-12的无血清培养上清进行浓缩,用阴离子交换色谱柱,阳离子交换色谱柱和尺寸排阻色谱柱纯化出rhIL-12;并对rhIL-12的纯度、活性及氨基酸序列进行检测。结果纯化后产物经Western blot鉴定(P35、P40)为rhIL-12,毛细管电泳法测得其纯度在98%以上;生物学活性达8.0×106IU/mg,且氨基酸序列与已报道的序列相一致。结论建立了获得高纯度、高生物活性的rhIL-12的纯化与鉴定方法。Purpose To explore the identification and purification methods of recombinmant human inter-leukin-12（rhIL-12）. Methods The Millipore ultrafiltration standard system was used to concentrate rhIL-12, then anion exchange chromatographic column, cation exchange chromatographic column and size exclusive chromatographic column were used in turn to purify the target protein. Meanwhile, the purity, bio-activity and amino acid sequencing of target protein were tested. Results The product of purification was identified as rhIL- 12 by detecting P35 and P40 using Western blot. The protein purity which was identified by the capillary elec-trophoresis was above 98%. The biology activity was 8.0×10^6 IU/mg. The amino acids sequence was consistent with the predicted sequence. Conclusion The purification method that could obtain rhIL-12 owning high purification and biology activity has been established successfully.

关 键 词：重组人白细胞介素12 纯化 鉴定 

分 类 号：R392-33[医药卫生—免疫学]