重组人前列腺特异性抗原纯化与活性鉴定  

Primary purification and activity assay of recombinant prostate-specific antigen

作　　者：邹冬辉[1] 张家颖[2] 路英丽[3] 郭焱[1] 王忠山[1] 

机构地区：[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

出　　处：《吉林大学学报（医学版）》2008年第1期1-5,共5页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(39870313)

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3.5h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。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.

关 键 词：前列腺特异性抗原 酶活性检测 分离纯化 鉴定 

分 类 号：Q786[生物学—分子生物学]