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Antibodies 2018, 7(4), 36; https://doi.org/10.3390/antib7040036

Genetic Fusion of an Anti-BclA Single-Domain Antibody with Beta Galactosidase

1
Naval Research Laboratory, Center for Biomolecular Science and Engineering, Washington, DC 20375, USA
2
The Washington Center for Internships and Academic Seminars, 1333 16th Street N.W., Washington, DC 20036, USA
3
Nova Research Inc., Alexandria, VA 22308, USA
*
Author to whom correspondence should be addressed.
Received: 12 September 2018 / Revised: 26 September 2018 / Accepted: 27 September 2018 / Published: 29 September 2018
(This article belongs to the Special Issue Nanobody)
Full-Text   |   PDF [2064 KB, uploaded 29 September 2018]   |  

Abstract

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 (β-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 β-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. View Full-Text
Keywords: Bacillus anthracis; immunoassay; single-domain antibody; genetic fusion; Beta galactosidase Bacillus anthracis; immunoassay; single-domain antibody; genetic fusion; Beta galactosidase
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Anderson, G.P.; Shriver-Lake, L.C.; Walper, S.A.; Ashford, L.; Zabetakis, D.; Liu, J.L.; Breger, J.C.; Brozozog Lee, P.A.; Goldman, E.R. Genetic Fusion of an Anti-BclA Single-Domain Antibody with Beta Galactosidase. Antibodies 2018, 7, 36.

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