猪血中高效提纯纤溶酶原方法的建立及其功能验证

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中图分类号：TS201.25 文献标志码：A 文章编号：0366-6964（2026）01-0454-10

Abstract： The purpose of this study is to establish an efficient method for purifying plasminogen from pig blood. In this study，sPLG was purified from pig blood using lysine affinity chromatography combined with size exclusion chromatography. Meanwhile，recombinant swine tissue plasminogen activator（st-PA） and staphylokinase（SAK）were expressed in Escherichia coli. Then，st-PA was purified by refolding of inclusion bodies，and SAK was purified by nickel column affinity chromatography combined with size exclusion chromatography. The activities of the prepared sPLG， SAK and st-PA were evaluated by a plasmin-specific substrate hydrolysis assay，a fibronectin degradation test，and an in vitro pig blood clot dissolution assay. The results showed that sPLG was highly purified by the method established by this study. Approximately 50mg of pure sPLG was obtained from 250mL of pig blood plasma. The purified sPLG could be activated by st-PA or SAK， and the resulting plasmin could then cleave the plasmin-specific substrate，degrade fibronectin，and lyse pig blood clots in uitro.In conclusion，this study established an efficient method for the preparation of sPLG， st-PA and SAK. In addition，the complete plasminogen activity of sPLG was verified using st-PA and SAK as well. This study provide essential materials for research on the mechanisms by which pathogens invade pigs via sPLG.

Keywords： plasminogen; tissue plasminogen activators; staphylokinase ; thrombus； protein purification Corresponding authors：FENG Zhixin，E-mail： fzxjaas@163. com；CHEN Rong，E-mail： chenronggrape@163.com

血栓，俗称血凝块，是由纤维蛋白交联成网格状并包裹红细胞和血小板等成分产生的块状物[1]，可以抑制血液流动。（剩余16456字）