Novel application of an in vitro technique to the detection and quantification of botulinum neurotoxin antibodies

• Published in: Journal of immunological methods 2004-05, Vol.288 (1), p.55-60
• Main Authors: Hall, Yper H.J, Chaddock, John A, Moulsdale, Hilary J, Kirby, Elizabeth R, Alexander, Frances C.G, Marks, James D, Foster, Keith A
• Format: Article
• Language: English
• Subjects: Animals; Anti-toxin; Antibodies - analysis; Antibodies - immunology; Antiserum; Biological and medical sciences; Botulinum neurotoxin; Botulinum Toxins - immunology; Botulinum Toxins - pharmacology; Clostridium botulinum; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Glycine - drug effects; Glycine - metabolism; Molecular immunology; Neurons - drug effects; Neurons - immunology; Rats; Rats, Sprague-Dawley; Spinal Cord - drug effects; Spinal Cord - immunology; Spinal cord culture; Techniques
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/j.jim.2004.02.011

Detection of Clostridium botulinum neurotoxin (BoNT) neutralising antibodies is currently achieved using the mouse lethality assay (MLA). This technique has provided the majority of the data for vaccine development and, with the increasing use of BoNT as a therapeutic agent, the MLA is the assay of choice to evaluate ‘non-responder’ antisera. However, the MLA is semi-quantitative and has an animal consumption rate that raises ethical concerns. The development of an alternative is therefore desirable. Here, we describe an in vitro neuronal release assay that may represent such an alternative in terms of both its sensitivity and ability to produce quantitative data. Initially recognised in the course of assessing a novel vaccine candidate, the suitability of this assay has been further explored using an International standard. The results support the conclusion that the detection of neutralising antibodies in human sera should be attempted using this method.