A new peptide substrate for enhanced botulinum neurotoxin type B detection by endopeptidase-liquid chromatography-tandem mass spectrometry/multiple reaction monitoring assay

作者：Rosen Osnat; Feldberg Liron; Gura Sigalit; Zichel Ran^*

来源：Analytical Biochemistry, 2015, 473: 7-10.

DOI：10.1016/j.ab.2014.09.016

• Botulinum neurotoxin
• Detection
• UPLC
• Mass spectrometry
• Peptide substrate

Botulinum neurotoxins (BoNTs) are the most toxic proteins in nature. Rapid and sensitive detection of BoNTs is achieved by the endopeptidase-mass spectrometry (Endopep-MS) assay. In this assay, BoNT cleaves a specific peptide substrate and the cleaved products are analyzed by MS. Here we describe the design of a new peptide substrate for improved detection of BoNT type B (BoNT/B) in the Endopep-MS assay. Our strategy was based on reported BoNT/B-substrate interactions integrated with analysis method efficiency considerations. Incorporation of the new peptide led to a 5-fold increased sensitivity of the assay both in buffer and in a clinically relevant human spiked serum.

• 出版日期2015-3-15