Serodiagnosis of cancer using porcine carboxypeptidase A as an animal antigen recognized by human monoclonal antibody HB4C5.
Protein from hog which is recognized by human monoclonal antibody (HB4C5), generated from a patient with large cell lung carcinoma, was identified as carboxypeptidase A by comparison of the protein with carboxypeptidase A in enzymatic activity, immunologic reactivity, and amino acid sequence. Carboxypeptidase A activity was also found in human cancer tissue, and purified antigen from cancer tissue recognized by the antibody HB4C5 was reacted with rabbit anti-carboxypeptidase A serum, indicating that carboxypeptidase A is an antigen of HB4C5. Since large amounts of carboxypeptidase A can be obtained from porcine sources, a simple method for its purification was established. The fraction which was most reactive with HB4C5 was obtained from acetone powder of porcine pancreas by successive applications of water extraction, ammonium sulfate precipitation, trypsin treatment, and Mono Q column chromatography. Its apparent molecular weight was 40,000, according to SDS polyacrylamide gel electrophoresis. When the reactivity of IgG in sera with the purified carboxypeptidase A was measured, the detection rates for lung, ovary, larynx, uterus, and liver cancer were more than 50%, while the rates for stomach and breast cancer were around 30%, and pancreatic cancer, benign diseases, and normal controls were minimally detected.
Hashizume
S
S
Morinaga Institute of Biological Science, Yokohama, Japan.
Mochizuki
K
K
Kamei
M
M
Kuroda
K
K
Kato
M
M
Sato
S
S
Yasumoto
K
K
Nakahashi
H
H
Tsuchimoto
K
K
Muraoka
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor / Q000032:analysis; D002268:Carboxypeptidases / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D043422:Carboxypeptidases A; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008175:Lung Neoplasms / Q000201:enzymology; D009369:Neoplasms / Q000175:diagnosis*; D010179:Pancreas / Q000201:enzymology; D012698:Serologic Tests; D013552:Swine
D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D002268:Carboxypeptidases; D043422:Carboxypeptidases A