用亲和层析法纯化人精浆蛋白

Purification of human γ seminoprotein with affinity chromatography

【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.