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Molecular characterization of human trk proto-oncogene product monoclonal antibodies
1Bristol-Myers Squibb Pharmaceutical Research Institute, Department of Molecular Biology, Princeton, New Jersey 08543-4000.
Abstract:
Monoclonal antibodies specific for the human trk protooncogene product, a tyrosine kinase receptor, have been produced from mice in which tumors were generated by injection of stably transfected NIH3T3 cells expressing the human trk proto-oncogene product. The panel of eleven antibodies are reactive in ELISA, immunostaining and immunoprecipitation. These antibodies bind to the extracellular domain of the human trk proto-oncogene product and demonstrate no cross-reactivity to the trk oncogene or murine trkB gene products. The antibodies are equally effective in recognizing human proto-trk when expressed by transfected human or mouse cell lines. These antibodies either bind to the carbohydrate moieties or are dependent upon the conformational structure created by the extensive glycosylations. The localization and nature of the epitopes recognized by these monoclonal antibodies were defined by immunoprecipitation analyses using several different trk oncoproteins. Potential applications of these antibodies are discussed.
Insights
Researchers developed novel monoclonal antibodies targeting the human trk proto-oncogene product, a tyrosine kinase receptor. These antibodies show high specificity and potential for various applications in cancer research.
Area of Science:
- Molecular Biology
- Immunology
- Oncology
Background:
- The trk proto-oncogene product is a tyrosine kinase receptor implicated in various cellular processes.
- Developing specific antibodies against human trk is crucial for understanding its role in oncogenesis.
Purpose of the Study:
- To generate and characterize a panel of monoclonal antibodies specific for the human trk proto-oncogene product.
- To investigate the binding characteristics and epitope recognition of these novel antibodies.
Main Methods:
- Production of monoclonal antibodies in mice immunized with NIH3T3 cells expressing human trk.
- Assays including ELISA, immunostaining, and immunoprecipitation to evaluate antibody reactivity and specificity.
- Epitope mapping using immunoprecipitation with various trk oncoproteins.
Main Results:
- Eleven monoclonal antibodies were successfully generated, reactive in ELISA, immunostaining, and immunoprecipitation.
- Antibodies specifically bind to the extracellular domain of human trk, with no cross-reactivity to trk oncogene or murine trkB.
- Antibodies recognize human trk expressed in both human and mouse cell lines, binding to carbohydrate moieties or conformation-dependent epitopes.
Conclusions:
- A valuable panel of specific monoclonal antibodies against the human trk proto-oncogene product has been developed.
- These antibodies offer potential applications in diagnostics and therapeutics for trk-related conditions.