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Rapid Testing for Botulinum Toxin using Egg Yolk Antibodies

Award No.: 
N-00014-04-1-0659
Status: 
Complete
Principal Investigator: 
Mark Cook
PI Organization: 
University of Wisconsin-Madison
Abstract: 
The objective of this research project was to demonstrate the capacity of the egg antibody platform in detection and neutralization of botulinum toxin. Anti-botulinum toxin antibody was produced by immunizing hens with botulinum toxoid in Freund complete adjuvant followed by several booster immunizations containing toxoid and increasing amounts of toxin in Freund incomplete adjuvant. Egg yolks were extracted with polyethylene glycol for antibody detection and neutralization experiments. Using chicken antibody, an affordable detection assay was developed capable of detecting toxoid at 1 pg/ml. In addition, chicken antibody binding to toxin was comparable to commercially available antibody. In a mouse toxin neutralization assay, antibody was incubated for 1 hour with dilutions of toxin, and 1 mg neutralized 4,000 intraperitoneal injected lethal doses of botulinum toxin. In one month, one hen can produce 260 liters of antibody detection reagents, or enough antibody to neutralize 11.6 million mouse lethal doses of botulinum toxin. In addition, antibody storage methods have been defined. Lyonphilized egg yolk antibody stored for 1 year at room temperature.
Analytic Methods Utilized: 
Academic Disciplines: