Two step procedure for purification of enzymatically active prostate-specific antigen from seminal plasma.
The role of prostate-specific antigen (PSA) during the onset of prostate cancer and subsequent tumor growth and metastasis is not well understood. We have developed a simple two step procedure, based on principles of hydrophobic charge-induction chromatography and molecular size chromatography to provide pure free-PSA (f-PSA) preparation that is free from all other known PSA complexes as well as human kallikrein 2 (hK2). The overall recovery of f-PSA is 72%. The isolated f-PSA consists of three known isoforms that corresponds to pI of 6.2, 6.4 and 7.2. f-PSA is enzymatically active and its enzymatic activity can be effectively neutralized by a serine protease inhibitor.
Bindukumar
B
B
Department of Molecular and Cellular Biology, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY 14263, USA.
Kawinski
Elzbieta
E
Cherrin
Craig
C
Gambino
Leah M
LM
Nair
Madhavan P N
MP
Schwartz
Stanley A
SA
Chadha
Kailash C
KC
D015153:Blotting, Western; D002850:Chromatography, Gel; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008297:Male; D017430:Prostate-Specific Antigen / Q000737:chemistry / Q000302:isolation & purification*; D012661:Semen / Q000276:immunology*
D017430:Prostate-Specific Antigen