Characterization of novel murine monoclonal antibodies directed against the extracellular domain of human HER2 tyrosine kinase receptor
| Autor: | Hodjatallah Rabbani, Mahmood Jeddi-Tehrani, Jalal Khoshnoodi, Fathollah Tahmasebi, Fazel Shokri, Tohid Kazemi, Neda Mohajer, Ali Ahmad Bayat |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
medicine.drug_class
Receptor ErbB-2 Immunology Blotting Western Enzyme-Linked Immunosorbent Assay Biology Monoclonal antibody Antibodies Monoclonal Humanized Proto-Oncogene Mas Epitope Receptor tyrosine kinase law.invention Antibodies Monoclonal Murine-Derived Mice law Antibody Specificity Cell Line Tumor medicine Extracellular Immunology and Allergy Animals Humans skin and connective tissue diseases neoplasms Mice Inbred BALB C Titrimetry Trastuzumab Flow Cytometry Molecular biology Recombinant Proteins Protein Structure Tertiary Cell culture Monoclonal biology.protein Recombinant DNA NIH 3T3 Cells Female Antibody Protein Binding |
| Zdroj: | Hybridoma (2005). 30(4) |
| ISSN: | 1557-8348 |
| Popis: | HER2 proto-oncogene encodes a transmembrane receptor tyrosine kinase overexpressed in a variety of solid tumors. Several mouse monoclonal antibodies (MAbs) have been developed that recognize the extracellular part of HER2; of them two MAbs were humanized and employed for targeted immunotherapy. In this study we aimed to produce murine MAbs that specifically recognize the extracellular domain of human HER2. BALB/c mice were first primed with HER2-transfected NIH-3T3 cells and then boosted with recombinant extracellular part of HER2. Splenocytes from hyperimmunized mice were fused with myeloma cells and growing hybridomas were selected and screened for HER2 reactivity by an indirect ELISA. HER2-specific hybridomas were selected, cloned by limiting dilution assay, and further characterized by Western blotting and flow cytometry techniques. All clones showed positive reactivity to HER2 with binding affinity, ranging from 1.9×10(8) to 5×10(9), and stained HER2-transfected cells and malignant cells overexpressing HER2. None of the MAbs inhibited the binding of trastuzumab (Herceptin(®)) to HER2, indicating recognition of distinct epitopes by these MAbs. Based on these findings, our MAbs could be potentially used for selective targeting of HER2-expressing malignancies. |
| Databáze: | OpenAIRE |
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