Derivation and Characterization of Monoclonal Antibodies Against Human Folypolyglutamate Synthetase
| Autor: | Shuang Luo, Steven H. Zuckerman, John F. Carpenter, Dotzlaf Joe Edward, Mcclure Don B, Deborah L. Fisher, Yue-Wei Qian, Xiliang Wang, Aimin Lin, Victor J. Chen, Mariam Ehsani, Rebecca R. Miles |
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| Rok vydání: | 2007 |
| Předmět: |
Protein Denaturation
Protein Conformation medicine.drug_class Molecular Sequence Data Immunology Monoclonal antibody Epitope Mice Antibody Specificity Neoplasms medicine Animals Humans Immunology and Allergy Amino Acid Sequence Peptide Synthases RNA Small Interfering Antiserum Mice Inbred BALB C Messenger RNA Hybridomas biology Antibodies Monoclonal Surface Plasmon Resonance Molecular biology Peptide Fragments Blot Epitope mapping Biochemistry Polyclonal antibodies Immunoglobulin G biology.protein RNA Interference Antibody Epitope Mapping HeLa Cells |
| Zdroj: | Hybridoma. 26:155-161 |
| ISSN: | 1557-8348 1554-0014 |
| Popis: | Folypolyglutamate synthetase (FPGS) plays a critical role in the cellular retention of both folates and antifolates. Resistance to antifolates is in part related to changes in FPGS enzyme activity and levels of messenger RNA, or in some instances, protein as evaluated by Western blots using polyclonal antisera. The present study was designed to derive a series of monoclonal antibodies (MAb) against the native protein, to characterize them in terms of specificity and epitope mapping, and to determine kinetic constants by Biacore. We report on 3 IgG(1) kappa MAbs-namely, 4-2, 4-3, and 4-18-with epitopes localized to the carboxyl domain of the protein. These antibodies recognize a single band on Western blots of HeLa cell lysates, which is significantly reduced following RNAi knockdown. The recognition of both the native and denatured conformations of FPGS by these MAbs should provide useful reagents for FPGS quantitation in either tumor cell lysates or in tumor biopsies. |
| Databáze: | OpenAIRE |
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