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Ex vivo assay evaluates botulinum toxin potency in mice, suggesting a humane alternative to traditional methods.
Botulinum neurotoxin (BoNT) is the most toxic substance known to mankind. Its mechanism of action is based on inhibition of acetylcholine release from nerve terminals. Traditionally, BoNT's potency has been measured by a mouse bioassay (MBA) controlling BoNT's binding, internalisation and intracellular cleavage function. BoNT’s potency is given in mouse units (MU, LD 50 units) defined as the amount of BoNT killing 50% of a specified mouse population following intraperitoneal injection. The mouse diaphragm assay (MDA) is an ex vivo assay measuring the twitch force of a mouse hemidiaphragm preparation after electric phrenic nerve stimulation. It was adapted by Hans Bigalke to measure BoNT potency. It drastically reduces animal consumption and agony, providing similar sensitivity in less time. It has also been used widely to detect and quantify neutralising BoNT antibodies.
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Tacik et al. (2026) studied this question.
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