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Proceedings Paper

Using llama derived single domain antibodies to target botulinum neurotoxins
Author(s): Marla D. Swain; George P. Anderson; Rachael D. Bernstein; Jinny L. Liu; Ellen R. Goldman
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Paper Abstract

Llama serum contains both conventional IgG as well as unique forms of antibody that contain only heavy chains where antigen binding is mediated through a single variable domain. These variable domains can be expressed recombinantly and are referred to as single domain antibodies (sdAb). SdAb are among the smallest known naturally derived antigen binding fragments, possess good solubility, thermal stability, and can refold after heat and chemical denaturation. Llamas were immunized with either BoNT A or B toxoid and phage display libraries prepared. Single domain antibodies (sdAb) that were able to detect botulinum neurotoxin (BoNT) serotypes A and B were selected from their respective libraries. Here, the binders obtained by panning the BoNT B library on either BoNT B toxoid or BoNT B complex toxoid coated plates or BoNT B toxin coupled microspheres are described. Using these panning methods, we selected for binders that showed specificity for BoNT B. Phage displayed binders were screened, moved to a protein expression vector and soluble sdAb was produced. Using a Luminex flow cytometer binders were evaluated in direct binding assays. We have exploited the unique properties of sdAb and used them as biological recognition elements in immuno-based sensors that can detect BoNT B.

Paper Details

Date Published: 23 April 2010
PDF: 9 pages
Proc. SPIE 7674, Smart Biomedical and Physiological Sensor Technologies VII, 767405 (23 April 2010); doi: 10.1117/12.852674
Show Author Affiliations
Marla D. Swain, U.S. Naval Research Lab. (United States)
George P. Anderson, U.S. Naval Research Lab. (United States)
Rachael D. Bernstein, Nova Research, Inc. (United States)
Jinny L. Liu, U.S. Naval Research Lab. (United States)
Ellen R. Goldman, U.S. Naval Research Lab. (United States)


Published in SPIE Proceedings Vol. 7674:
Smart Biomedical and Physiological Sensor Technologies VII
Brian M. Cullum; D. Marshall Porterfield; Karl S. Booksh, Editor(s)

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