Characterization of monoclonal antibodies for prostate-specific antigen and development of highly sensitive free prostate-specific antigen assays.
BACKGROUND
The recent elucidation of the importance of serological free prostate-specific antigen (PSA) in the diagnosis of prostate cancer has created a demand for immunoassays specific for free PSA.


METHODS
We developed and characterized 11 monoclonal antibodies with high affinities for PSA (Ka values from 1.1 x 10(8) to 1.8 x 10(10)L/mol), only 3 of which cross-react with human glandular kallikrein (hK2). Using these antibodies and PSA antibodies developed by others, in conjunction with time-resolved fluorometry, we developed ultrasensitive sandwich immunoassays specific for the free form of PSA.


RESULTS
The analytical detection limit of these immunoassays is 0.001 microg/L. To our knowledge, this is the most sensitive free PSA assay reported to date. The free PSA immunoassays exhibit <1% cross-reactivity with PSA-alpha1-antichymotrypsin, show no cross-reactivity with hK2, and correlate well with established free PSA kits. The 11 antibodies developed by our group, in conjunction with 4 commercially available antibodies, were used to generate a putative epitope map of the PSA molecule.


CONCLUSIONS
The highly sensitive free PSA immunoassays may be used for measuring PSA subfractions in female serum, an application currently impossible with other reported free PSA immunoassays.
Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, 600 University Avenue, Toronto, Ontario, Canada M5G 1X5.
M H
MH
Black
C L
CL
Grass
J
J
Leinonen
U H
UH
Stenman
E P
EP
Diamandis
D000818:Animals; D000911:Antibodies, Monoclonal; D003429:Cross Reactions; D000939:Epitopes; D015200:Fluoroimmunoassay / Q000379:methods; D006801:Humans; D007610:Kallikreins / Q000032:analysis / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D017430:Prostate-Specific Antigen / Q000097:blood* / Q000276:immunology; D011933:Reagent Kits, Diagnostic; D020840:Tissue Kallikreins
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011933:Reagent Kits, Diagnostic; D007610:Kallikreins; D020840:Tissue Kallikreins; D017430:Prostate-Specific Antigen