用亲和层析法纯化人精浆蛋白  

Purification of human γ seminoprotein with affinity chromatography

作　　者：孙脊峰[1] 杨洁[1] 郝晓柯[2] 金明[3] 赵晶[3] 王成济[3] 杨安钢[3] 

机构地区：[1]第四军医大学唐都医院肾脏内科,陕西西安710038 [2]西京医院临床分子生物学研究中心,陕西西安710032 [3]第四军医大学生物化学与分子生物学教研室,陕西西安710032

出　　处：《武警医学院学报》2006年第2期91-92,169,共3页Acta Academiae Medicinae CPAPF

基　　金：国家"863"高技术计划课题资助项目(2001AA215321);教育部高等学校骨干教师资助项目(2000-65-66);国家杰出青年科学基金资助项目(39925036)

摘　　要：【目的】用亲和层析法从增生的前列腺组织中纯化人精浆蛋白(-γseminoprotein,-γsm)。【方法】采用超声波粉碎、NP-40提取与两种免疫亲和层析相结合[抗人精浆蛋白单抗(E4B7mAb)亲和层析和蛋白A亲和层析]的新提纯方案,获得了高纯度-γsm抗原。【结果】所纯化的-γsm蛋白经ELISA双抗夹心法和免疫印迹分析检测证明,用此法从增生的前列腺组织中分离到-γsm抗原,而且纯化的-γsm具有生物活性。【结论】这种方法的建立将为分离-γsm抗原提供一种有效途径,为抗精浆蛋白基因工程抗体的研究提供良好的抗原保证。[Objective] To purify human γ seminoprotein （γ-sm） from hyperplastic prostate tissue by affinity chromatography. [Methods] Highly purified γ-sin antigen was obtained by ultrasonic disintegration, extracted with NP-40 and two kinds of affinity chromatographies of E4BTmAb-CNBr-Sepharose 4B and Protein A-Sepharose 4B. [Results] Purified γ-sm was detected by ELISA double antibodies sandwich and Western blot. The results showed that γ-sm proteins could be purified from hyperplastic prostate tissue and purified γ-sm had better biological activity. [Condusion] This result provides an effective way of purifying γ-sm antigen and lays a good antigen foundation for the research of engineering antibody against γ-sm.

关 键 词：亲和层析 纯化 人精浆蛋白 

分 类 号：R737.25[医药卫生—肿瘤]