Botulinum neurotoxin type A (BoNT A) is one of the most potent toxins to humans and has become an imminent threat in biological warfare. Administered as either a pre- or post-exposure prophylaxis, high affinity neutralizing antibodies can potentially provide protection against the disease botulism. Human single-chain variable fragment antibodies (scFvs) that recognize BoNT A have previously been identified, including one antibody that can almost completely neutralize BoNT A. While extremely promising, it would be beneficial to enhance the affinity of these antibodies towards BoNT A. This will be achieved by subjecting the scFVs to affinity maturation/molecular evolution methods in combination with phage-display. The most promising antibodies will then be converted to a more stable format for further characterization. The ability of the antibodies to protect against the cytotoxic effects of BoNT A will be determined with an in vitro cell-based assay. Additionally, binding affinity and specificity of these antibodies will be investigated using BIAcore. The association rate constant, dissociation rate constant, and equilibrium dissociation constant for antibody-BoNT A binding will be obtained for the evolved antibodies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32AI062014-03
Application #
7076173
Study Section
Special Emphasis Panel (ZRG1-F07 (20))
Program Officer
Van de Verg, Lillian L
Project Start
2004-06-16
Project End
2007-06-15
Budget Start
2006-06-16
Budget End
2007-06-15
Support Year
3
Fiscal Year
2006
Total Cost
$52,048
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
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Eubanks, Lisa M; Hixon, Mark S; Jin, Wei et al. (2007) An in vitro and in vivo disconnect uncovered through high-throughput identification of botulinum neurotoxin A antagonists. Proc Natl Acad Sci U S A 104:2602-7
Eubanks, Lisa M; Dickerson, Tobin J; Janda, Kim D (2007) Technological advancements for the detection of and protection against biological and chemical warfare agents. Chem Soc Rev 36:458-70
Brogan, Andrew P; Eubanks, Lisa M; Koob, George F et al. (2007) Antibody-catalyzed oxidation of delta(9)-tetrahydrocannabinol. J Am Chem Soc 129:3698-702
Eubanks, Lisa M; Rogers, Claude J; Beuscher 4th, Albert E et al. (2006) A molecular link between the active component of marijuana and Alzheimer's disease pathology. Mol Pharm 3:773-7
Eubanks, Lisa M; Dickerson, Tobin J; Janda, Kim D (2005) Vitamin B2-mediated cellular photoinhibition of botulinum neurotoxin A. FEBS Lett 579:5361-4