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Ricin Detection Using Phage Displayed Single Domain Antibodies
Naval Research Laboratory, Center for Bio/Molecular Science and Engineering, 4555 Overlook Ave SW, Washington DC 20375, USA
Nova Research Inc, 1900 Elkin Street, Suite 230, Alexandria VA 22308 USA
Clark Atlanta University, Department of Biological Sciences, Atlanta GA 30314 USA
* Author to whom correspondence should be addressed.
Received: 31 October 2008; in revised form: 13 January 2009 / Accepted: 14 January 2009 / Published: 19 January 2009
Abstract: Phage-displayed single domain antibodies (sdAb) were compared to monomeric solubly expressed sdAb and llama polyclonal antibodies for the detection of ricin. SdAb are comprised of the variable domain derived from camelid heavy chain only antibodies (HcAb). Although HcAb lack variable light chains, they as well as their derivative sdAb are able to bind antigens with high affinity. The small size of sdAb (~16 kDa), while advantageous in many respects, limits the number of labels that can be incorporated. The ability to incorporate multiple labels is a beneficial attribute for reporter elements. Opportunely, sdAb are often selected using phage display methodology. Using sdAb displayed on bacteriophage M13 as the reporter element gives the potential for incorporating a very high number of labels. We have demonstrated the use of both sdAb and phage- displayed sdAb for the detection of ricin using both enzyme linked immunosorbent assays (ELISAs) and Luminex fluid array assays. The phage-displayed sdAb led to five to ten fold better detection of ricin in both the ELISA and Luminex assays, resulting in limits of detection of 1 ng/mL and 64 pg/mL respectively. The phage-displayed sdAb were also dramatically more effective for the visualization of binding to target in nitrocellulose dot blot assays, a method frequently used for epitope mapping.
Keywords: Ricin; single domain antibody; phage; Luminex; phage display
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Cite This Article
MDPI and ACS Style
Goldman, E.R.; Liu, J.L.; Bernstein, R.D.; Swain, M.D.; Mitchell, S.Q.; Anderson, G.P. Ricin Detection Using Phage Displayed Single Domain Antibodies. Sensors 2009, 9, 542-555.
Goldman ER, Liu JL, Bernstein RD, Swain MD, Mitchell SQ, Anderson GP. Ricin Detection Using Phage Displayed Single Domain Antibodies. Sensors. 2009; 9(1):542-555.
Goldman, Ellen R.; Liu, Jinny L.; Bernstein, Rachael D.; Swain, Marla D.; Mitchell, Stanley Q.; Anderson, George P. 2009. "Ricin Detection Using Phage Displayed Single Domain Antibodies." Sensors 9, no. 1: 542-555.