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.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
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)
关键词
前列腺特异性抗原
酶活性检测
分离纯化
鉴定
prostate specific antigen
activity assay
purification
identification
