Abstract
Botulinum neurotoxins (BoNTs) are very potent toxins and category A biological threat agents. BoNT serotypes /C1 and /D affect birds and mammals and can be potentially lethal to humans. We have previously described the usefulness of the Endopep-MS method to detect the activity of BoNT A through G. This report was followed by the application of the method to clinical samples. The activity of the BoNT serotypes associated with human disease (/A, /B, /E, and /F) was successfully detected. However, BoNT/C and /D require different conditions for fast substrate cleavage, and a comprehensive description of a method to study BoNT/C and /D has not yet been reported. This work describes a new, optimized version of the Endopep-MS method to detect BoNTs /C1 and /DC either spiked directly in 20μL of reaction buffer or spiked in a larger volume of buffer and further extracted using antibody-coated magnetic beads. It was found that the incubation temperature at 42°C was more effective for both toxin serotypes, but each toxin serotype has an optimum cleavage pH. Additionally, we describe for the first time a proteomics study using a fast trypsin digestion method and label-free quantification of these toxin serotypes. FEMS Immunology & Medical Microbiology
Original language | English (US) |
---|---|
Pages (from-to) | 288-300 |
Number of pages | 13 |
Journal | FEMS Immunology and Medical Microbiology |
Volume | 61 |
Issue number | 3 |
DOIs | |
State | Published - Apr 2011 |
Externally published | Yes |
Keywords
- Botulism
- Endopep-MS
- Label-free quantification
- MS
- Proteomics
ASJC Scopus subject areas
- Immunology and Allergy
- Microbiology
- Immunology
- Microbiology (medical)
- Infectious Diseases