重组人B淋巴细胞刺激因子纯化及其生物活性鉴定

Purification and bioactivity identification of recombinant human B-lymphocyte stimulator

目的 获得高纯度和高活性的B淋巴细胞刺激因子。方法 重组原核表达载体pQE 80L BLyS112 -2 85在大肠杆菌DH5α中经IPTG诱导表达rhBLyS112 -2 85,超声碎菌 ,提取包涵体 ,经Ni2 + NTA亲和层析和SepharcrylS 2 0 0凝胶过滤层析纯化后 ,选择不同氧化还原条件进行复性 ,进而检测其生物学活性。结果 得到了有较高纯度和较好生物学活性的rhBLyS112 -2 85蛋白。结论 优化了BLyS蛋白的纯化和复性的参数 。

Objective To prepare highly purified and refolded B-lymphocyte stimulator. Methods The expression of rhBLyS 112-285 in E.coli was induced by IPTG, and the bacteria were split by sonication.The rhBLyS 112-285 was purified by Ni 2+-NTA resin affinity chromato- graphy and Sepharcryl S-200 gel filtration chromatography. The purified protein was refolded under different conditions. Then the bioactivity of the protein was analyzed. Results The rhBLyS 112-285 with high purity and bioactivity was attained. Conclusion The parameters of purification and refolding of BLyS were optimized, which may pave a way for further studies.