Analysis of monoclonal antibodies to prostate-specific antigen: reactivity with native and recombinant prostate-specific antigen.
Epitope mapping analysis was performed on 53 antibodies submitted to the ISOBM TD-3 PSA Workshop. Western blotting and N-terminal amino acid sequencing, using both native and recombinant human prostate-specific antigen (rPSA), identified four different epitope groups for native PSA. Under reducing conditions native PSA was not recognized by 18/53 antibodies suggesting they reacted with conformation-dependent epitopes. Nine other antibodies reacted with a rPSA polypeptide doublet of 34-35 kD corresponding to different rPSA glycoforms. From sequence mapping studies 22/53 antibodies bound epitopes within amino acid residues 25-85, 3/53 antibodies bound to epitopes within residues 86-220, while 10/53 antibodies bound epitopes within residues 221-261. These results indicate that there are multiple immunogenic epitopes localized on the PSA molecule.
Zhou
Z
Z
Bayer Corporation, Business Group Diagnostics, Tarrytown, NY 10591, USA.
Barnett
T R
TR
Very
D L
DL
Ng
P C
PC
Pellegrino
C M
CM
Davis
G
G
Belenky
A
A
Allard
W J
WJ
Yeung
K K
KK
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D016367:Baculoviridae / Q000378:metabolism; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D018604:Epitope Mapping; D006801:Humans; D008297:Male; D011467:Prostate / Q000276:immunology; D017430:Prostate-Specific Antigen / Q000235:genetics / Q000276:immunology*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012661:Semen / Q000276:immunology; D020539:Sequence Analysis, Protein
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D011994:Recombinant Proteins; D017430:Prostate-Specific Antigen