Catalytic antibodies for complex reactions |
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Authors: | Binodh S DeSilva György Orosz Kamal L Egodage Robert G Carlson Richard L Schowen George S Wilson |
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Institution: | 1. Procter and Gamble Pharmaceuticals, Rt. 320, Norwich, NY 2. Department of Organic Chemistry, E?tv?s Loránd University, Budapest 3. Monsanto Co., 800 North Lindbergh Blvd., St. Louis, MO 4. Department of Chemistry, University of Kansas, 66045, Lawrence, KS
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Abstract: | Success in generating catalytic antibodies as enzyme mimics lies in the strategic design of the transition-state analog (TSA)
for the reaction of interest, and careful development of screening processes for the selection of antibodies that are catalysts.
Typically, the choice of TSA structure is straightforward, and the criterion for selection in screening is often binding of
the TSA to the antibody in a microtiter-plate assay. This article emphasizes the problems of TSA design in complex reactions
and the importance of selecting antibodies on the basis of catalysis as well as binding to the TSA. The target reaction is
the derivatization of primary amines with naphthalene-2,3-dicarboxaldehyde (NDA) in the presence of cyanideion. The desired
outcome is selective catalysis of formation of the fluorescent derivative in preference to nonfluorescent side-products. In
the study, TSA design was directed toward the reaction branch leading to the fluorescent product. Here, we describe a microtiter
plate-based assay that is capable of detecting antibodies showing catalytic activity atan early stage. Of the antibodies selected,
36% showed no appreciable binding to any of the substrates tested, but did show catalytic activity in deriving one or more
of the amino acids screened. In contrast, only two out of 77 clones that showed binding did not show catalysis. Thus, in this
complex system, observation of binding is a good predictor of the presence of catalytic activity, and failure to observe binding
is a poor predictor of the absence of catalytic activity. |
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Keywords: | Catalytic antibody transition-state analog NDA/CN reaction screening enzyme mimics |
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