Acevedo Boris, Perera Yasser, Torres Edel, Pentón David, Ayala Marta, Gavilondo Jorge
Center for Genetic Engineering and Biotechnology, Havana, Cuba.
Prostate. 2006 Jul 1;66(10):1029-36. doi: 10.1002/pros.20267.
Prostate specific antigen (PSA) is a relevant antigen in diagnosis; follow-up, and therapeutic approaches for fighting the prostate cancer. Several methods have been published previously to obtain a high purity preparation of PSA. In general, these methods are expensive, time-consuming, laborious, and in some cases produce low yields.
Based on a panel of 7 anti-PSA Mab's we carried on binding and elution experiments of PSA antigen in 96-well plates. The selected Mab were immobilized in a Sepharose CL-4B activated matrix with the purpose of purify PSA from human seminal fluid. In order to optimize the purification procedure, we test several washing and elution conditions (chaotropic agents, high ionic strength solution, and extreme pH).
We selected a high ionic strength solution (2 M MgCl2) as elution condition, and a previous washing step with a mix of two ionic solutions (2.5 M NaCl pH 8/1 M MgCl2 pH 5.5) in order to purify PSA. Using such conditions we obtained a PSA preparation with 90% of purity and 50% of recovery.
In this article, we report a simple, quickly, and non-expensive procedure to obtain free-PSA from human seminal plasma at high purity levels.
前列腺特异性抗原（PSA）是前列腺癌诊断、随访及治疗方法中的一种相关抗原。此前已发表了多种获取高纯度PSA制剂的方法。总体而言，这些方法昂贵、耗时、费力，且在某些情况下产率较低。
基于一组7种抗PSA单克隆抗体，我们在96孔板中进行了PSA抗原的结合和洗脱实验。将所选单克隆抗体固定在活化的琼脂糖CL-4B基质上，目的是从人精液中纯化PSA。为了优化纯化程序，我们测试了几种洗涤和洗脱条件（离液剂、高离子强度溶液和极端pH值）。
我们选择高离子强度溶液（2 M MgCl2）作为洗脱条件，并在洗脱前用两种离子溶液（2.5 M NaCl pH 8/1 M MgCl2 pH 5.5）的混合液进行洗涤步骤以纯化PSA。使用这些条件，我们获得了纯度为90%、回收率为50%的PSA制剂。
在本文中，我们报告了一种简单、快速且廉价的方法，可从人精浆中获得高纯度的游离PSA。
