The test consists of differentiated human neuroblastoma (SiMa) in combination with a sandwich ELISA procedure. The sensitivity is comparable to in vivo test in mice. The ELISA measures the increase of protein SNAP-25 cleavage depending on the botulinum toxin A bioactivity.

Botulismustoxin prevents the fusion of acetylcholine-filled vesicles with the plasma membrane of excitatory synapses cleaving the resident SNAP-25 proteins. Therefore acetylcholine (neurotransmitter) cannot released released into the synaptic cleft and the stimulus of a synapse can not be passed to the next. This ultimately leads to a relaxation of the muscles.

The new method is used in order to test the stability and potency of Allergans´ products BOTOX ® (onabotulinumtoxinA) and BOTOX ® Cosmetic. Until then, mice had to suffer from respiratory muscle paralysis and death in countless required tests. With the new procedure Allergan can save up to 95% of animal testing. The in vitro test is only applicable for Allergan products of botulinum type A. The company has invested about 65 million U.S. dollars for the test research and development.

Applied locally the BOTOX ® from Allergan is a product for a variety of applications such as for the treatment of cervical dystonia, muscle stiffness or spasticity as well as certain forms of chronic migraine. The same recipe is used for cosmetic purposes in wrinkle treatment. The effect lasts for about 3 months, then the toxin will be degraded. Than the organism produces again fully functional fusion proteins (SNAP-25).

After approval by the U.S. FDA and the EU, in this year´s february the company got green light from the German Federal Institute for Drugs and Medical Products (BfArM) for the use of the in vitro test.

European manufacturers are going to develop their own in-vitro tests for its own Botox products. They want to adopt this test or their own test developments specifically to their products. However, there are any obstacles to overcome: in April the Frankfurt manufactorer Merz reported about setbacks in the development of such a test method.

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