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Establishing a Method for Efficient Purification of Plasminogen from Swine Blood and Verifying Its Function
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Ke MA1, 2, Lei ZHANG2, 3, 4, Fei HAO2, 3, Xing XIE2, 3, Zhenzhen ZHANG2, 3, Guoqing SHAO2, 3, Rong CHEN2, 3, Zhixin FENG1, 2, 3, 4
Acta Veterinaria et Zootechnica Sinica | 2026, 57(1) : 454 - 463
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Acta Veterinaria et Zootechnica Sinica | 2026, 57(1): 454-463
• BASIC VETERINARY MEDICINE •
Establishing a Method for Efficient Purification of Plasminogen from Swine Blood and Verifying Its Function
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Ke MA1, 2, Lei ZHANG2, 3, 4, Fei HAO2, 3, Xing XIE2, 3, Zhenzhen ZHANG2, 3, Guoqing SHAO2, 3, Rong CHEN2, 3, Zhixin FENG1, 2, 3, 4
Affiliations
  • 1.School of Life Sciences，Nanjing Agricultural University，Nanjing 210095，China
  • 2.Key Laboratory of Veterinary Biological Products Engineering，Ministry of Agriculture and Rural Affairs，Institute of Veterinary Medicine，Jiangsu Academy of Agricultural Sciences，Nanjing 210014，China
  • 3.Guotai Technology Innovation Center of Veterinary Biological Products （Taizhou），Taizhou 225300，China
  • 4.College of Animal Medicine，Nanjing Agricultural University，Nanjing 210095，China
Published: 2026-01-23 doi: 10.11843/j.issn.0366-6964.2026.01.040
Outline
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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 50 mg of pure sPLG was obtained from 250 mL 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 vitro. 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.

plasminogen  /  tissue plasminogen activators  /  staphylokinase  /  thrombus  /  protein purification
马可, 张磊, 郝飞, 谢星, 张珍珍, 邵国青, 陈蓉, 冯志新. 猪血中高效提纯纤溶酶原方法的建立及其功能验证. 畜牧兽医学报, 2026 , 57 (1) : 454 -463 . DOI: 10.11843/j.issn.0366-6964.2026.01.040
Ke MA, Lei ZHANG, Fei HAO, Xing XIE, Zhenzhen ZHANG, Guoqing SHAO, Rong CHEN, Zhixin FENG. Establishing a Method for Efficient Purification of Plasminogen from Swine Blood and Verifying Its Function[J]. Acta Veterinaria et Zootechnica Sinica, 2026 , 57 (1) : 454 -463 . DOI: 10.11843/j.issn.0366-6964.2026.01.040
Appendix
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Year 2026 volume 57 Issue 1
141
61
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Article Info
doi: 10.11843/j.issn.0366-6964.2026.01.040
  • Receive Date：2024-11-18
  • Online Date：2026-09-16
  • Published：2026-01-23
Article Data
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History
  • Received：2024-11-18
Funding
Affiliations
    1.School of Life Sciences，Nanjing Agricultural University，Nanjing 210095，China
    2.Key Laboratory of Veterinary Biological Products Engineering，Ministry of Agriculture and Rural Affairs，Institute of Veterinary Medicine，Jiangsu Academy of Agricultural Sciences，Nanjing 210014，China
    3.Guotai Technology Innovation Center of Veterinary Biological Products （Taizhou），Taizhou 225300，China
    4.College of Animal Medicine，Nanjing Agricultural University，Nanjing 210095，China

Corresponding:

FENG Zhixin，E-mail：
CHEN Rong， E-mail：
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表12种不同金属材料的力学参数
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Family		 属数
Number of
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Number of
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Percentage of
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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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