Autor: |
Adva Mechaly, Einat B. Vitner, Yinon Levy, David Gur, Moria Barlev-Gross, Assa Sittner, Michal Koren, Haim Levy, Emanuelle Mamroud, Morly Fisher |
Jazyk: |
angličtina |
Rok vydání: |
2021 |
Předmět: |
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Zdroj: |
Pathogens, Vol 10, Iss 3, p 285 (2021) |
Druh dokumentu: |
article |
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: |
Directory of Open Access Journals |
Externí odkaz: |
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