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Proceedings Paper

SdAb heterodimer formation using leucine zippers
Author(s): Ellen R. Goldman; George P. Anderson; P. Audrey Brozozog-Lee; Dan Zabetakis
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Paper Abstract

Single domain antibodies (sdAb) are variable domains cloned from camel, llama, or shark heavy chain only antibodies, and are among the smallest known naturally derived antigen binding fragments. SdAb derived from immunized llamas are able to bind antigens with high affinity, and most are capable of refolding after heat or chemical denaturation to bind antigen again. We hypothesized that the ability to produce heterodimeric sdAb would enable reagents with the robust characteristics of component sdAb, but with dramatically improved overall affinity through increased avidity. Previously we had constructed multimeric sdAb by genetically linking sdAb that bind non-overlapping epitopes on the toxin, ricin. In this work we explored a more flexible approach; the construction of multivalent binding reagents using multimerization domains. We expressed anti-ricin sdAb that recognize different epitopes on the toxin as fusions with differently charged leucine zippers. When the initially produced homodimers are mixed the leucine zipper domains will pair to produce heterodimers. We used fluorescence resonance energy transfer to confirm heterodimer formation. Surface plasmon resonance, circular dichroism, enzyme linked immunosorbent assays, and fluid array assays were used to characterize the multimer constructs, and evaluate their utility in toxin detection.

Paper Details

Date Published: 29 May 2013
PDF: 9 pages
Proc. SPIE 8723, Sensing Technologies for Global Health, Military Medicine, and Environmental Monitoring III, 872313 (29 May 2013); doi: 10.1117/12.2016145
Show Author Affiliations
Ellen R. Goldman, U.S. Naval Research Lab. (United States)
George P. Anderson, U.S. Naval Research Lab. (United States)
P. Audrey Brozozog-Lee, Nova Research, Inc. (United States)
Dan Zabetakis, U.S. Naval Research Lab. (United States)

Published in SPIE Proceedings Vol. 8723:
Sensing Technologies for Global Health, Military Medicine, and Environmental Monitoring III
Šárka O. Southern, Editor(s)

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