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目的 :为了从细胞株培养液中纯化长半衰期的t PA组合突变体FrGGI。方法 :采用高表达克隆细胞株 ,进行大量培养 ,收集 2 4h细胞培养液为初始材料 ,利用t PAK2区含有Lysine结合位点的特性和Zn2 +与蛋白质中咪唑、巯基等基团螯合作用 ,经Lysine Sepharose 4B亲和层析和Zn2 + Sepharose 4B金属螯合层析二步组合纯化。结果 :获得高比活的FrGGI,为研究该突变体生物学特性及推广应用奠定基础。结论 :利用赖氨酸和锌离子柱二步组合纯化方法 ,可有效地分离纯化t PA及其含K2区的突变体
OBJECTIVE: In order to purify long-lived t PA combination mutant FrGGI from cell line culture. Methods: Highly expressing clonal cell lines were cultured for 24 h. The cell culture medium was collected for 24 h. Using t PAK2 region containing Lysine binding site and Zn2 + chelation with imidazole and sulfhydryl groups in protein, Purified by Lysine Sepharose 4B affinity chromatography and Zn2 + Sepharose 4B metal chelate chromatography. Results: The FrGGI with high specific activity was obtained, which laid the foundation for the study on the biological characteristics of the mutant and its application. Conclusion: Two-step purification of lysine and zinc ion column can effectively separate and purify t PA and its K2-containing mutants