Toxins 2010, 2(8), 2198-2212; doi:10.3390/toxins2082198
Article

Characterization of SNARE Cleavage Products Generated by Formulated Botulinum Neurotoxin Type-A Drug Products

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Received: 9 July 2010; in revised form: 10 July 2010 / Accepted: 18 August 2010 / Published: 19 August 2010
(This article belongs to the Special Issue Toxins as Therapeutics)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: The study evaluated substrate cleavage product(s) generated by three botulinum neurotoxin serotype A (BoNT/A) medicinal drug products utilizing a novel and highly specific, light-chain activity, high-performance liquid chromatography (LCA-HPLC) method. Samples were reacted with a commercially available BoNT/A fluorescent substrate derived from the SNAP-25 sequence. Reaction products were separated by reversed-phase HPLC. The method detected an atypical cleavage pattern by one of the formulated drug products. IncobotulinumtoxinA produced two cleavage fragments rather than the single fragment typically generated by BoNT/A. Identification confirmed the secondary cleavage at a position corresponding to SNAP-25 Arg198–Ala199 (normal BoNT/A cleavage is Gln197–Arg198). Arg198–Ala199 is also the cleavage site for trypsin and serotype C toxin. Normal cleavage was observed for all other BoNT/A drug product samples, as well as 900-kD and 150-kD bulk toxin BoNT/A. The reason for this unexpected secondary cleavage pattern by one formulated BoNT/A drug product is unknown. Possible explanations include a contaminating protease and/or damage to the 150-kD type-A toxin causing nonspecific substrate recognition and subsequent cleavage uncharacteristic of type-A toxin. The BoNT/A drug products were also analyzed via the LCA-HPLC assay using a commercial BoNT/C fluorescent substrate derived from the syntaxin sequence. Cleavage of the serotype C substrate by incobotulinumtoxinA was also confirmed whilst neither of the other drug products cleaved the syntaxin substrate.
Keywords: abotulinumtoxinA; botulinum toxin; BOTOX®; Dysport®; incobotulinumtoxinA; onabotulinumtoxin A; SNAP-25; SNARE; syntaxin; Xeomin®
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MDPI and ACS Style

Hunt, T.; Rupp, D.; Shimizu, G.; Tam, K.; Weidler, J.; Xie, J. Characterization of SNARE Cleavage Products Generated by Formulated Botulinum Neurotoxin Type-A Drug Products. Toxins 2010, 2, 2198-2212.

AMA Style

Hunt T, Rupp D, Shimizu G, Tam K, Weidler J, Xie J. Characterization of SNARE Cleavage Products Generated by Formulated Botulinum Neurotoxin Type-A Drug Products. Toxins. 2010; 2(8):2198-2212.

Chicago/Turabian Style

Hunt, Terrence; Rupp, David; Shimizu, Gary; Tam, Karen; Weidler, Julia; Xie, Jack. 2010. "Characterization of SNARE Cleavage Products Generated by Formulated Botulinum Neurotoxin Type-A Drug Products." Toxins 2, no. 8: 2198-2212.

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