Conventional Test Method:

Mouse LD₅₀ Assay for Botulinum Toxin Potency Testing

Alternative Test Method:

Name & Description:

In Vitro Cell-Based Assay

The release of neurotransmitter from embryonic spinal cord neurons after 16 h exposure to BoNT/A, and the reduction in the inhibition of release when antibodies neutralize BoNT/A, are measured.

Validation Status:

US: ICCVAM workshop panel found that no cell-based method could currently be used to reduce or replace animals for BoNT potency testing or for detecting botulinum toxin in environmental or biological samples. In their current state of development, cell-based methods are not sufficiently sensitive. Which cell types and/or cell lines are the most appropriate is also not known (2008).

Regulatory Status:

None

Effect or Potential Effect on Animal Use:

Replacement

References:

• Adler S., Bicker G., Bigalke H., Bishop C., Blümel J., Dressler D., Fitzgerald J., Gessler F., Heuschen H., Kegel B., Luch A., Milne C., Pickett A., Ratsch H., Ruhdel I., Sesardic D., Stephens M., Stiens G., Thornton P.D., Thürmer R., Vey M., Spielmann H., Grune B. and Liebsch M. (2010) The current scientific and legal status of alternative methods to the LD50 test for Botulinum neurotoxin potency testing. ATLA 38(4):315-330.
• Hall Y.H., Chaddock J.A., Moulsdale H.J., Kirby E.R., Alexander F.C., Marks J.D. and Foster K.A. (2004) Novel application of an in vitro technique to the detection and quantification of botulinum neurotoxin antibodies. Journal of Immunological Methods 288(1-2):55-60.
• ICCVAM (2008) Report on the ICCVAM-NICEATM/ECVAM Scientific Workshop on Alternative Methods to Refine, Reduce or Replace the Mouse LD₅₀ Assay for Botulinum Toxin Testing. Bethesda MD: National Institutes of Health (NIH).

Information Last Reviewed:

May 2012