Distribution and physical properties of BCA200, a Mr 200,000 glycoprotein selectively associated with human breast cancer.
Of 122 mouse monoclonal antibodies selective for human breast cancer, 13 immunoprecipitated an acidic glycoprotein from SK-Br-3 and ZR-75-30 human breast cancer cells. The antigen (BCA200) migrates with an apparent molecular weight of 200,000 on reducing and 180,000 on nonreducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting a single polypeptide chain with a folded domain stabilized by a disulfide bond. Cross-blocking and sandwich immunoassays detected at least three distinct antigenic determinants on BCA200. Scatchard experiments measured 1,000,000 to 5,000,000 antigen copies per SK-Br-3 cell. The tissue distribution of BCA200 was studied using two monoclonals to different epitopes. Neither antibody stained any cells in human blood. When frozen sections of 20 normal human tissues were immunoperoxidase stained, the only positive structures were mucinous glands of colon, transitional epithelium of bladder, sweat glands of skin, and acinar epithelium of breast. Antibody 454C11 stained 16 of 21 breast tumor frozen sections and 9 of 12 breast cancer cell lines, while antibody 520C9 stained 5 of 20 breast tumors and 4 of 10 breast cancer lines. Cross-reaction was observed with lung, prostatic, pancreatic, endometrial, and ovarian cancer, but not with lymphoma, melanoma, colon, stomach, bladder, or esophageal cancer. When conjugated to ricin A chain, 10 of 13 antibodies produced immunotoxins selectively cytotoxic to SK-Br-3 breast cancer cells.
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Cetus Corporation, Department of Immunology, Emeryville, California 94608.
Kassel
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JA
Hsieh-Ma
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Bjorn
M J
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Tringale
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F
Eaton
A M
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Reid
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Frankel
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Nadji
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D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000032:analysis*; D001940:Breast / Q000276:immunology; D001943:Breast Neoplasms / Q000276:immunology*; D015180:Electrophoresis, Gel, Two-Dimensional; D000939:Epitopes / Q000276:immunology; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D008970:Molecular Weight; D011233:Precipitin Tests
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000939:Epitopes; D006023:Glycoproteins; C059645:breast cancer-associated antigen 200, human