Genetic Fusion of an Anti-BclA Single-Domain Antibody with Beta Galactosidase
| Autor: | Lauryn Ashford, Lisa C. Shriver-Lake, George M. Anderson, Scott A. Walper, Jinny L. Liu, Joyce C. Breger, Dan Zabetakis, P. Audrey Brozozog Lee, Ellen R. Goldman |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
lcsh:Immunologic diseases. Allergy Phage display 030106 microbiology Immunology genetic fusion Article 03 medical and health sciences Drug Discovery medicine Immunology and Allergy Beta-galactosidase immunoassay Bacillus anthracis single-domain antibody biology medicine.diagnostic_test Beta galactosidase Chemistry fungi Protein engineering biology.organism_classification 030104 developmental biology Single-domain antibody Biochemistry Immunoassay biology.protein Antibody lcsh:RC581-607 Function (biology) |
| Zdroj: | Antibodies, Vol 7, Iss 4, p 36 (2018) Antibodies Volume 7 Issue 4 |
| ISSN: | 2073-4468 |
| Popis: | The Bacillus collagen-like protein of anthracis (BclA), found in Bacillus anthracis spores, is an attractive target for immunoassays. Previously, using phage display we had selected llama-derived single-domain antibodies that bound to B. anthracis spore proteins including BclA. Single-domain antibodies (sdAbs), the recombinantly expressed heavy domains from the unique heavy-chain-only antibodies found in camelids, provide stable and well-expressed binding elements with excellent affinity. In addition, sdAbs offer the important advantage that they can be tailored for specific applications through protein engineering. A fusion of a BclA targeting sdAb with the enzyme Beta galactosidase (&beta gal) would enable highly sensitive immunoassays with no need for a secondary reagent. First, we evaluated five anti-BclA sdAbs, including four that had been previously identified but not characterized. Each was tested to determine its binding affinity, melting temperature, producibility, and ability to function as both capture and reporter in sandwich assays for BclA. The sdAb with the best combination of properties was constructed as a fusion with &beta gal and shown to enable sensitive detection. This fusion has the potential to be incorporated into highly sensitive assays for the detection of anthrax spores. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|