This proposal presents an immunotherapeutic strategy for the treatment of cocaine addiction in which catalytic antibodies are evolved for their ability to hydrolyze cocaine to its non-psychoactive products, ecognine methyl ester and benzoic acid. Using previously identified cocaine catalysts, termed GNL, as a starting point, two different directed evolution techniques will be applied for the construction of novel scFv libraries. The first involves affinity maturation by complementarity determining region (CDR) walking based on unpublished crystal structure results from the Wilson and Janda laboratories, while the second relies on random recombination, or deoxyribose nucleic acid (DMA) shuffling. These GNLM (GNL-mutant) libraries will be screened using high throughput methodology and assessed for their ability to hydrolyze cocaine; the top two antibodies will be determined by a comparison their kinetic parameters. Selected GNLM catalysts will be examined for their ability to affect locomotor activity in rats, since motor behavior provides a sensitive and reproducible measure of drug action under standardized conditions.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
1F32DA018024-01A1
Application #
6884432
Study Section
Human Development Research Subcommittee (NIDA)
Program Officer
Babecki, Beth
Project Start
2005-04-01
Project End
2007-03-31
Budget Start
2005-04-01
Budget End
2006-03-31
Support Year
1
Fiscal Year
2005
Total Cost
$47,296
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
McKenzie, Kathleen M; Mee, Jenny M; Rogers, Claude J et al. (2007) Identification and characterization of single chain anti-cocaine catalytic antibodies. J Mol Biol 365:722-31
Dickerson, Tobin J; McKenzie, Kathleen M; Hoyt, Amanda S et al. (2007) Phage escape libraries for checkmate analysis. Proc Natl Acad Sci U S A 104:12703-8