Abstract |
The molecular nature of an osteosarcoma-associated antigen was investigated with the three monoclonal antibodies Ost6 ( immunoglobulin ( IgG1), Ost7 ( IgG1), and Ost15 ( IgG2a), which selectively react with frozen sections of osteosarcoma and chondrosarcoma tissues. When tested with a panel of 41 human cell lines in the mixed hemadsorption assay, the antibodies reacted similarly with three of six osteosarcomas, one choriocarcinoma, one teratoma, and one osteoblast-like culture, but failed to react with 32 lines of normal and other tumor cell types. Immunoprecipitation plus sodium dodecyl sulfate (SDS)-- polyacrylamide gel electrophoresis and sequential immunoprecipitation studies revealed that in [35S] methionine- or [14C] glucosamine-labelled osteosarcoma cells the three antibodies detected a single glycoprotein, with an apparent molecular mass of 86 kilodaltons (kDa), which was not affected by reducing conditions. Tunicamycin treatment and pulse-chase experiments showed glycosylation of this molecule to be N-linked; it arose from a 54-kDa polypeptide precursor. Alkaline phosphatase activity was detected in the material rich in 86-kDa molecules that was immunoprecipitated from serologically reactive cell lines with each antibody. These antibodies also cross-reacted with two isoenzymes of alkaline phosphatase (strongly with the liver and bone, and moderately with the placental isoenzyme), but not with the intestinal form.
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Authors | T Nakamura, M Gross, T Yamamuro, S K Liao |
Journal | Biochemistry and cell biology = Biochimie et biologie cellulaire
(Biochem Cell Biol)
Vol. 65
Issue 12
Pg. 1091-100
(Dec 1987)
ISSN: 0829-8211 [Print] Canada |
PMID | 3331286
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Antibodies, Monoclonal
- Antigens, Neoplasm
- Glycoproteins
- Alkaline Phosphatase
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Topics |
- Alkaline Phosphatase
(analysis)
- Antibodies, Monoclonal
- Antigens, Neoplasm
(analysis)
- Autoradiography
- Binding, Competitive
- Cross Reactions
- Electrophoresis, Polyacrylamide Gel
- Glycoproteins
(analysis)
- Glycosylation
- Hemadsorption
- Humans
- Immunologic Techniques
- Osteosarcoma
(diagnosis, immunology)
- Tumor Cells, Cultured
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