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Fast and novel purification method to obtain the prostate specific antigen (PSA) from human seminal plasma
Author(s) -
Acevedo Boris,
Perera Yasser,
Torres Edel,
Pentón David,
Ayala Marta,
Gavilondo Jorge
Publication year - 2006
Publication title -
the prostate
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.295
H-Index - 123
eISSN - 1097-0045
pISSN - 0270-4137
DOI - 10.1002/pros.20267
Subject(s) - prostate specific antigen , elution , chromatography , chaotropic agent , ionic strength , antigen , prostate cancer , chemistry , sepharose , prostate , matrix (chemical analysis) , cancer , immunology , biochemistry , medicine , aqueous solution , enzyme
Background 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. Methods 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). Results We selected a high ionic strength solution (2 M MgCl 2 ) as elution condition, and a previous washing step with a mix of two ionic solutions (2.5 M NaCl pH 8/1 M MgCl 2 pH 5.5) in order to purify PSA. Using such conditions we obtained a PSA preparation with 90% of purity and 50% of recovery. Conclusion In this article, we report a simple, quickly, and non‐expensive procedure to obtain free‐PSA from human seminal plasma at high purity levels. Prostate 66: 1029–1036, 2006. © 2006 Wiley‐Liss, Inc.