Biomol Ther.  2020 Jul;28(4):302-310. 10.4062/biomolther.2019.200.

Alternative Methods for Testing Botulinum Toxin: Current Status and Future Perspectives

  • 1College of Pharmacy, Yeungnam University, Gyeongsan 38541, Republic of Korea


Botulinum toxins are neurotoxic modular proteins composed of a heavy chain and a light chain connected by a disulfide bond and are produced by Clostridium botulinum. Although lethally toxic, botulinum toxin in low doses is clinically effective in numerous medical conditions, including muscle spasticity, strabismus, hyperactive urinary bladder, excessive sweating, and migraine. Globally, several companies are now producing products containing botulinum toxin for medical and cosmetic purposes, including the reduction of facial wrinkles. To test the efficacy and toxicity of botulinum toxin, animal tests have been solely and widely used, resulting in the inevitable sacrifice of hundreds of animals. Hence, alternative methods are urgently required to replace animals in botulinum toxin testing. Here, the various alternative methods developed to test the toxicity and efficacy of botulinum toxins have been briefly reviewed and future perspectives have been detailed.


Botulinum toxin; Acetylcholine; In vitro; Alternative studies
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