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Linking Single Domain Antibodies that Recognize Different Epitopes on the Same Target
Nova Research Inc., 1900 Elkin Street, Suite 230, Alexandria, VA 22308, USA
Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, 4555 Overlook Ave. SW, Washington, DC 20375, USA
* Author to whom correspondence should be addressed.
Received: 13 December 2011; in revised form: 13 January 2012 / Accepted: 29 January 2012 / Published: 1 February 2012
Abstract: Single domain antibodies (sdAb) are the recombinantly expressed variable regions from the heavy-chain-only antibodies found in camelids and sharks. SdAb are able to bind antigens with high affinity, and most are capable of refolding after heat or chemical denaturation to bind antigen again. Starting with our previously isolated ricin binding sdAb determined to bind to four non-overlapping epitopes, we constructed a series of sdAb pairs, which were genetically linked through peptides of different length. We designed the series so that the sdAb are linked in both orientations with respect to the joining peptide. We confirmed that each of the sdAb in the constructs was able to bind to the ricin target, and have evidence that they are both binding ricin simultaneously. Through this work we determined that the order of genetically linked sdAb seems more important than the linker length. The genetically linked sdAb allowed for improved ricin detection with better limits of detection than the best anti-ricin monoclonal we evaluated, however they were not able to refold as well as unlinked component sdAb.
Keywords: avidity; single domain antibody; ricin detection
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Cite This Article
MDPI and ACS Style
Glaven, R.H.; Anderson, G.P.; Zabetakis, D.; Liu, J.L.; Long, N.C.; Goldman, E.R. Linking Single Domain Antibodies that Recognize Different Epitopes on the Same Target. Biosensors 2012, 2, 43-56.
Glaven RH, Anderson GP, Zabetakis D, Liu JL, Long NC, Goldman ER. Linking Single Domain Antibodies that Recognize Different Epitopes on the Same Target. Biosensors. 2012; 2(1):43-56.
Glaven, Richard H.; Anderson, George P.; Zabetakis, Dan; Liu, Jinny L.; Long, Nina C.; Goldman, Ellen R. 2012. "Linking Single Domain Antibodies that Recognize Different Epitopes on the Same Target." Biosensors 2, no. 1: 43-56.