کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8553719 | 1562692 | 2018 | 9 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
In vitro potency determination of botulinum neurotoxin serotype A based on its receptor-binding and proteolytic characteristics
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کلمات کلیدی
GT1bH-chainNAPLD50PBSSV2CHCCATeNTL-chainBoNTTMAOBSA - BSAtris(2-carboxyethyl)phosphine - tris (2-carboxyethyl) فسفینIn vitro assay - آزمایش درون آزمایشگاهیbovine serum albumin - آلبومین سرم گاوtetanus neurotoxin - ترومبوس نوروتوکسینActivity determination - تعیین فعالیتlight chain - زنجیره سبکheavy chain - زنجیره سنگینTCEP - ساکتPhosphate-buffered saline - محلول نمک فسفات با خاصیت بافریBotulinum neurotoxin - نوروتوکسین بوتولینوم
موضوعات مرتبط
علوم زیستی و بیوفناوری
علوم محیط زیست
بهداشت، سم شناسی و جهش زایی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Botulinum neurotoxins (BoNTs) inhibit the release of the neurotransmitter acetylcholine from motor neurons, resulting in highly effective muscle relaxation. In clinical and aesthetic medicine, serotype BoNT/A, which is most potent for humans, is widely used to treat a continuously increasing spectrum of disorders associated with muscle overactivity. Because of the high toxicity associated with BoNTs, it is mandatory to precisely determine the potency of every batch produced for pharmaceutical purposes. Here we report a new quantitative functional in vitro assay for BoNT/A. In this binding and cleavage (BINACLE) assay, the toxin is first bound to specific receptor molecules. Then a chemical reduction is performed, thereby releasing the light chain of BoNT/A and activating its proteolytic domain. The activated light chain is finally exposed to its substrate protein SNAP-25, and the fragment resulting from the proteolytic cleavage of this protein is quantified in an antibody-mediated reaction. The BoNT/A BINACLE assay offers high specificity and sensitivity with a detection limit below 0.5 mouse lethal dose (LD50)/ml. In conclusion, this new in vitro assay for determining BoNT/A toxicity represents an alternative to the LD50 test in mice, which is the “gold standard” method for the potency testing of BoNT/A products.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Toxicology in Vitro - Volume 53, December 2018, Pages 80-88
Journal: Toxicology in Vitro - Volume 53, December 2018, Pages 80-88
نویسندگان
Heike A. Behrensdorf-Nicol, Emina Wild, Ursula Bonifas, Jolanta Klimek, Kay-Martin Hanschmann, Beate Krämer, Birgit Kegel,