Egg Yolk Antibodies for Detection and Neutralization of Clostridium botulinum Type A Neurotoxin

• Published in: Journal of food protection 2009-05, Vol.72 (5), p.1005-1011
• Main Authors: Trott, D.L, Yang, M, Gonzalez, J, Larson, A.E, Tepp, W.H, Johnson, E.A, Cook, M.E
• Format: Article
• Language: English
• Subjects: Animals; Antibodies; Antibodies, Monoclonal; antibody detection; Antibody Specificity; Bacteria; Bioassays; Biological & chemical terrorism; Biological and medical sciences; Biological Assay; Botulinum toxin; Botulinum Toxins, Type A - immunology; Botulinum Toxins, Type A - isolation & purification; botulism; Clostridium botulinum; Clostridium botulinum A; Egg production; Egg Yolk; Eggs; Enzyme-Linked Immunosorbent Assay; Enzymes; Food contamination & poisoning; Food Contamination - analysis; Food industries; Food Microbiology; food pathogens; Food safety; Freund's Adjuvant; Fundamental and applied biological sciences. Psychology; General aspects; Immunization; Immunoassay; Lethal Dose 50; Medical research; Methods of analysis, processing and quality control, regulation, standards; Mice; Neurotoxins; Neutralization; Neutralization Tests; Polyethylene glycol; Poultry; Rabbits; Reagents; Research projects; Toxicity; Toxins; toxoids
• ISSN: 0362-028X; 1944-9097
• DOI: 10.4315/0362-028X-72.5.1005

The objective of this research project was to determine the usefulness of an egg antibody platform for producing materials for the detection and neutralization of botulinum type A neurotoxin. Yield estimates for detection and neutralizing antibodies produced using methods described were calculated. Antibody specific to botulinum toxoid A (aToxoid) and toxin A (aBoNT/A) was produced by immunizing hens with botulinum toxoid A (toxoid) followed by increasing amounts of botulinum neurotoxin A (BoNT/A) in Freund incomplete adjuvant. Egg yolks were extracted with polyethylene glycol (PEG) for antibody detection and neutralization experiments. A model aToxoid/toxoid immunoassay using only egg yolk antibody was developed and had a detection limit of 1 pg/ml of toxoid. In an indirect enzyme-linked immunosorbent assay of BoNT/A-specific antibody, the aBoNT/A contained more BoNT/A-specific antibody than did the aToxoid, and aBoNT/A was as effective as commercial rabbit antibody. The aToxoid provided no protection against BoNT/A in a standard mouse neutralization assay; however, 1 mg of PEG-extracted aBoNT/A neutralized 4,000 lethal doses of BoNT/A injected intraperitoneally. Based on these results, we calculated that in 1 month one hen could produce more than 100 liters of antibody detection reagents or enough antibody to neutralize approximately 11.6 million mouse lethal doses of botulinum toxin. Utilization of an egg antibody platform is potentially rapid (28 to 70 days) and scalable to kilogram quantities using current egg production facilities with as few as 1,000 hens.