Fast and novel purification method to obtain the prostate specific antigen (PSA) from human seminal plasma.
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 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.


CONCLUSIONS
In this article, we report a simple, quickly, and non-expensive procedure to obtain free-PSA from human seminal plasma at high purity levels.
Acevedo
Boris
B
Center for Genetic Engineering and Biotechnology, Havana, Cuba. boris.acevedo@cigb.edu.cu
Perera
Yasser
Y
Torres
Edel
E
Pentón
David
D
Ayala
Marta
M
Gavilondo
Jorge
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000941:Antigens / Q000276:immunology; D002846:Chromatography, Affinity / Q000191:economics / Q000379:methods*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007118:Immunoassay / Q000191:economics / Q000379:methods*; D008024:Ligands; D008297:Male; D051379:Mice; D017430:Prostate-Specific Antigen / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D012661:Semen / Q000737:chemistry* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000941:Antigens; D008024:Ligands; D017430:Prostate-Specific Antigen