Characterization of anti-prostatic acid phosphatase monoclonal antibody and its medical significance.
Monoclonal antibody to purified prostatic acid phosphatase from seminal plasma was produced by fusion of spleen cells from immunized mice with the Sp2/O-Ag 14 cell line. This hybridoma-derived antibody, designated MAb-14, was classified as IgG1 immunoglobulin. The apparent affinity constant of phosphatase.MAb-14 complex formation calculated by using the Langmuir isotherm is 2.4 x 10(-8) M. The molecular weight of the complex formed under the condition of antibody excess was found to be 350K, which suggests that 2 molecules of prostatic phosphatase bind to 1 molecule of the antibody. The MAb-14 antibody bound to phosphatase had a negligible effect on the catalytic activity of the enzyme. All isoenzymatic forms of catalytically active prostatic phosphatase, resolved by isoelectric focusing or by chromatofocusing in different pH gradients, reacted with the monoclonal antibody. Several peptides of Mr 25K to 76K and of 13K to 76K were adsorbed from the prostatic tissue extract and from seminal fluid, respectively, on an MAb-14-Sepharose column. The MAb-14 monoclonal antibody was applied to the immunohistochemical investigation of prostatic phosphatase distribution in normal human prostate gland, in nodular hyperplasia, and in adenocarcinoma of the prostate. Immunostaining was observed in prostatic secretory epithelium, within the luminal content of prostatic glands, and in the neighborhood of prostatic cancer cells. Metastatic prostatic carcinoma was also strongly immunoreactive with the antibody. There was no cross-reactivity with leukocytes, kidney, liver, pancreas, spleen, breast, stomach mucosal, and colon tissues.
Kuciel
R
R
Institute of Medical Biochemistry, Copernicus Academy of Medicine, Kraków, Poland.
Mazurkiewicz
A
A
Ostrowski
W S
WS
Stachura
J
J
Steuden
I
I
Szkudlarek
J
J
Radzikowski
C
C
D000135:Acid Phosphatase / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal; D002384:Catalysis; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008970:Molecular Weight; D011467:Prostate / Q000201:enzymology*
D000911:Antibodies, Monoclonal; D000135:Acid Phosphatase