重组人白细胞介素12的纯化与鉴定

Identification and purification of recombinant human interleukin-12

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.