Epitope Binning of Novel Monoclonal Anti F1 and Anti LcrV Antibodies and Their Application in a Simple, Short, HTRF Test for Clinical Plague Detection
| Autor: | Moria Barlev-Gross, Haim Levy, Emanuelle Mamroud, Einat B. Vitner, Michal Koren, Morly Fisher, Yinon Levy, David Gur, Adva Mechaly, Assa Sittner |
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| Rok vydání: | 2021 |
| Předmět: |
Microbiology (medical)
medicine.drug_class lcsh:Medicine Monoclonal antibody Article Virulence factor Epitope 03 medical and health sciences Antigen Epitope binning medicine Immunology and Allergy LcrV Molecular Biology 030304 developmental biology 0303 health sciences General Immunology and Microbiology biology Chemistry lcsh:R 030302 biochemistry & molecular biology Molecular biology plague F1 Infectious Diseases Monoclonal biology.protein immuno-detection monoclonal antibodies Antibody HTRF |
| Zdroj: | Pathogens Volume 10 Issue 3 Pathogens, Vol 10, Iss 285, p 285 (2021) |
| ISSN: | 2076-0817 |
| DOI: | 10.3390/pathogens10030285 |
| Popis: | Mouse monoclonal antibodies were raised against plague disease biomarkers: the bacterial capsular protein fraction 1 (F1) and the low-calcium response—LcrV virulence factor (Vag). A novel tandem assay, employing BioLayer Interferometry (BLI), enabled the isolation of antibodies against four different epitopes on Vag. The tandem assay was carried out with hybridoma supernatants, circumventing the need for antibody purification. The BioLayer assay was further adopted for characterization of epitope-repetitive antigens, enabling the discovery of two unique epitopes on F1. The selected antibodies were purified and applied as “oligo-clonal” reagents for the immuno-detection of both biomarkers. The developed Homogenous Time Resolved Fluorescence (HTRF) tests were short (10 min) and simple (no washing steps), allowing for detection of 10 ng/mL F1 and 2.5 ng/mL Vag. The tests were successfully applied for detection of disease biomarkers produced by various Y. pestis strains during growth in blood culture vials. |
| Databáze: | OpenAIRE |
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