优化源自SNAP-25的肽底物，以改进内肽酶质谱分析法中肉毒杆菌A的检测。
Optimization of SNAP-25-derived peptide substrate for improved detection of botulinum A in the Endopep-MS assay.
作者信息
Rosen Osnat, Feldberg Liron, Dor Eyal, Gura Sigalit, Zichel Ran
机构信息
Department of Biotechnology, Israel Institute for Biological Research, Israel.
Department of Analytical Chemistry, Israel Institute for Biological Research, Israel.
出版信息
Anal Biochem. 2017 Jul 1;528:34-37. doi: 10.1016/j.ab.2017.04.012. Epub 2017 Apr 24.
Botulinum neurotoxins (BoNTs) are the most toxic proteins in nature. Endopeptidase-mass-spectrometry (Endopep-MS) is used as a specific and rapid in-vitro assay to detect BoNTs. In this assay, immunocaptured toxin cleaves a serotype-specific-peptide-substrate, and the cleavage products are then detected by MS. Here we describe the design of a new peptide substrate for improved detection of BoNT type A (BoNT/A). Our strategy was based on reported BoNT/A-SNAP-25 interactions integrated with analysis method efficiency considerations. Integration of the newly designed substrate led to a 10-fold increase in the assay sensitivity both in buffer and in clinically relevant samples.
肉毒杆菌神经毒素（BoNTs）是自然界中毒性最强的蛋白质。内肽酶质谱分析法（Endopep-MS）被用作一种特异性快速体外检测法来检测BoNTs。在该检测法中，免疫捕获的毒素切割一种血清型特异性肽底物，然后通过质谱法检测切割产物。在此，我们描述了一种新型肽底物的设计，用于改进A型肉毒杆菌神经毒素（BoNT/A）的检测。我们的策略基于已报道的BoNT/A与突触小体相关蛋白25（SNAP-25）的相互作用，并综合考虑了分析方法的效率。新设计底物的整合使缓冲液和临床相关样本中的检测灵敏度提高了10倍。
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