Quantitative differentiation of the haptoglobin-related gene product from haptoglobin in human plasma: a possible test for tumor-associated antigen.
The haptoglobin-related gene product (HGRP) having greater than 90% sequence homology with plasma haptoglobin, has recently been implicated as a tumor maker of recurrent breast cancer. Therefore, availability of a monoclonal antibody which is specific to the HGRP and ensuing ELISAs to quantitate the HGRP without cross-reactivity with plasma haptoglobin will allow determination of the clinical value of the HGRP in human plasma under various pathophysiological conditions. A peptide representing the first 34 residues of the N-terminal portion of the HRGP was synthesized and conjugated to keyhole limpet hemocyanin (KLH) to immunize Balb/c mice. Hybrid clones that produced specific antibodies to HRGP were screened with the same peptide but conjugated to bovine serum albumin (HRGP-BSA). From 12 hybridoma clones that recognized HGRP-peptide, we obtained one IgM producing clone (27.5) by limiting dilution technique that selectively reacted with HGRP-peptide with minimal cross-reactivity with haptoglobin. Using a separate monoclonal antibody, clone 21.7, which is directed to the alpha subunit of haptoglobin, we developed an enzyme-linked immunosorbent assay (ELISA) to determine the quantities of the HGRP in human plasma, which will allow assessment of HGRP as a clinical marker of malignancy.
Oh
S K
SK
Department of Microbiology, Boston University School of Medicine, MA 02118.
Kim
S H
SH
Abbruzzi
G
G
Adler
W H
WH
Tauber
A I
AI
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor / Q000097:blood; D001798:Blood Proteins / Q000276:immunology / Q000302:isolation & purification*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006041:Goats; D006242:Haptoglobins / Q000276:immunology / Q000302:isolation & purification*; D006433:Hemocyanins; D006801:Humans; D006825:Hybridomas; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008875:Middle Aged; D008974:Mollusca; D011817:Rabbits; D012710:Serum Albumin, Bovine / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D001798:Blood Proteins; C058632:HPR protein, human; D006242:Haptoglobins; D012710:Serum Albumin, Bovine; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin