SSB/La蛋白在HEK293F细胞中的高效表达、纯化及初步应用

Efficient expression,purification and preliminary application of SSB/La protein in HEK293F cells

目的天然SSB/La抗原存在难获取、成本高等问题,故使用HEK293F细胞尝试表达重组SSB/La蛋白。方法调取人SSB/La序列并进行基因密码子优化和合成,亚克隆至哺乳动物重组表达质粒后转染HEK293F细胞,完成培养后组合使用DEAE离子交换层析和Ni亲和层析纯化目的蛋白。结果重组SSB/La蛋白在HEK293F细胞中成功表达,随后纯化获得了高纯度的重组SSB/La蛋白,且进一步通过抗SSB抗体检测试剂盒初步证实了重组SSB/La蛋白具有优良的性能。结论成功实现了SSB/La蛋白制备,为其工业化生产及商业化应用奠定坚实的理论基础。

Objective The natural SSB/La antigen is difficult to obtain and expensive,the recombinant expression of SSB/La protein was attempted using HEK293F cells.Methods The human SSB/La sequences were acquired for gene codon optimization and synthesis,and then sub-cloned into mammalian recombinant vector and transfected into HEK293F cells.After cell culture,the target protein was purified by DEAE ion exchange chromatography and Ni affinity chromatography.Results The recombinant SSB/La protein was successfully expressed in HEK293F cells,and high-purity SSB/La protein was generated.Further,it was preliminary confirmed that the recombinant SSB/La protein had excellent properties through Autobio anti-SSB antibody test kit.Conclusion The preparation of SSB/La protein has been successfully achieved,laying solid theoretical foundations for its industrial production and commercial application.