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Antibodies 2017, 6(1), 3; doi:10.3390/antib6010003

Pairing Alpaca and Llama-Derived Single Domain Antibodies to Enhance Immunoassays for Ricin

1
US Naval Research Laboratory, Center for Biomolecular Science and Engineering, 4555 Overlook Ave SW, Washington, DC 20375, USA
2
US Naval Research Laboratory, HBCU Internship Program, 4555 Overlook Ave SW, Washington, DC 20375, USA
3
Nova Research Inc., 1900 Elkin St., Suite 230, Alexandria, VA 22308, USA
*
Authors to whom correspondence should be addressed.
Received: 18 October 2016 / Accepted: 17 January 2017 / Published: 2 February 2017
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Abstract

Previously, our group isolated and evaluated anti-ricin single domain antibodies (sdAbs) derived from llamas, engineered them to further increase their thermal stability, and utilized them for the development of sensitive immunoassays. In work focused on the development of therapeutics, Vance et al. 2013 described anti-ricin sdAbs derived from alpacas. Herein, we evaluated the utility of selected alpaca-derived anti-ricin sdAbs for detection applications, and engineered an alpaca-derived sdAb to increase its melting temperature, providing a highly thermal stable reagent for use in ricin detection. Four of the alpaca-derived anti-ricin A-chain sdAbs were produced and characterized. All four bound to epitopes that overlapped with our previously described llama sdAbs. One alpaca sdAb, F6, was found to possess both a high melting temperature (73 °C) and to work optimally with a thermally stable llama anti-ricin sdAb in sandwich assays for ricin detection. We employed a combination of consensus sequence mutagenesis and the addition of a non-canonical disulfide bond to further enhance the thermal stability of F6 to 85 °C. It is advantageous to have a choice of recognition reagents when developing assays. This work resulted in defining an additional pair of highly thermal stable sdAbs for the sensitive detection of ricin. View Full-Text
Keywords: camelid; llama; alpaca; single domain antibody; disulfide bond; thermal stability; ricin camelid; llama; alpaca; single domain antibody; disulfide bond; thermal stability; ricin
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MDPI and ACS Style

Turner, K.B.; Hardy, S.; Liu, J.L.; Zabetakis, D.; Lee, P.A.B.; Goldman, E.R.; Anderson, G.P. Pairing Alpaca and Llama-Derived Single Domain Antibodies to Enhance Immunoassays for Ricin. Antibodies 2017, 6, 3.

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