J4

• 基础研究 •    下一篇

重组人前列腺特异性抗原纯化与活性鉴定

邹冬辉1,张家颖2,路英丽3,郭 焱1,王忠山1   

  1. (1.吉林大学基础医学院细胞生物学教研室,吉林 长春 130021;2. 吉林大学基础医学院生物化学与分子生物学实验中心, 吉林 长春 130021;3.吉林大学第二医院妇产科, 吉林 长春130041)
  • 收稿日期:2007-08-18 修回日期:1900-01-01 出版日期:2008-01-28 发布日期:2008-01-28
  • 通讯作者: 王忠山

Primary purification and activity assay of recombinant prostate-specific antigen

ZOU Dong-hui1,ZHANG Jia-ying2,LU Ying-li3,GUO Yan1,WANG Zhong-shan1   

  1. (1.Department of Cell Biology,School of Basic Medical Sciences,Jilin University,Changchun 130021,China;2.Experimental Center of Biochemistry and Molecular Biology,School of Basic Medical Sciences,Jilin University,Changchun 130021,China;3.Department of Obstetrics and Gynecology,Second Hospital,Jilin University,Changchun 130041,China)
  • Received:2007-08-18 Revised:1900-01-01 Online:2008-01-28 Published:2008-01-28
  • Contact: WANG Zhong-shan

摘要: 目的:纯化和鉴定重组人前列腺特异性抗原(PSA),为PSA 的生物学特征研究、临床检测和治疗应用提供关键材料。方法: 采用阳离子交换层析技术纯化重组PSA(rPSA),利用Western blotting 鉴定其特异性,利用PSA底物S-2586检测rPSA活性。结果:最适甲醇诱导浓度为1.5%,经鉴定纯化的rPSA为人PSA。在不同反应条件下,在3.5 h内酶活性与时间成正比,rPSA 活性显著高于正常精浆PSA酶活性(P<0.05),酶活性的最适温度为30℃。结论:成功纯化出具有酶活性的rPSA。

关键词: 酶活性检测, 分离纯化, 鉴定

Abstract: Abstract:Objective To purify and assay the recombinant prostate-specific antigen (rPSA) to provide key material for study of PSA biology character,clinic detection and therapeutic application. Methods The rPSA was purified by cation-exchange chromatography from the yeast cell culture supernatant,then rPSA specificity was analyzed by Western blotting,and enzyme activity was assayed using s-2586 substrate. Results The most suitable methanol concentration to induce expression was 1.5%.The rPSA was purified by cation-exchange chromatography.The rPSA had chymotrypsin-like activity of PSA.Under different conditions enzyme activity was directly related with time.The most suitable temperature of enzyme activity was 30℃ within 3.5 h.The chymotrypsin-like activity of rPSA was obviously higher than that of PSA in seminal vesicle fluid of normal male. Conclusion The active,mature rPSA has been successfully purified.

Key words: activity assay, purification, identification

中图分类号: 

  • Q786