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Article

Targeted Mass Spectrometry Analysis of Clostridium perfringens Toxins

1
Faculty of Military Health Sciences, University of Defense in Brno, Třebešská 1575, CZ-500 01 Hradec Králové, Czech Republic
2
Department of Biology, Faculty of Science, University of Hradec Kralove, Hradecká 1285, CZ-500 03 Hradec Kralove, Czech Republic
3
Military Health Institute, Military Medical Agency, Tychonova 1, CZ-160 00 Prague 6, Czech Republic
*
Author to whom correspondence should be addressed.
Received: 1 March 2019 / Revised: 18 March 2019 / Accepted: 21 March 2019 / Published: 23 March 2019
Targeted proteomics recently proved to be a technique for the detection and absolute quantification of proteins not easily accessible to classical bottom-up approaches. Due to this, it has been considered as a high fidelity tool to detect potential warfare agents in wide spread kinds of biological and environmental matrices. Clostridium perfringens toxins are considered to be potential biological weapons, especially the epsilon toxin which belongs to a group of the most powerful bacterial toxins. Here, the development of a target mass spectrometry method for the detection of C. perfringens protein toxins (alpha, beta, beta2, epsilon, iota) is described. A high-resolution mass spectrometer with a quadrupole-Orbitrap system operating in target acquisition mode (parallel reaction monitoring) was utilized. Because of the lack of commercial protein toxin standards recombinant toxins were prepared within Escherichia coli. The analysis was performed using proteotypic peptides as the target compounds together with their isotopically labeled synthetic analogues as internal standards. Calibration curves were calculated for each peptide in concentrations ranging from 0.635 to 1101 fmol/μL. Limits of detection and quantification were determined for each peptide in blank matrices. View Full-Text
Keywords: mass spectrometry; PRM; Clostridium perfringens; protein toxin; epsilon toxin mass spectrometry; PRM; Clostridium perfringens; protein toxin; epsilon toxin
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MDPI and ACS Style

Duracova, M.; Klimentova, J.; Myslivcova Fucikova, A.; Zidkova, L.; Sheshko, V.; Rehulkova, H.; Dresler, J.; Krocova, Z. Targeted Mass Spectrometry Analysis of Clostridium perfringens Toxins. Toxins 2019, 11, 177. https://0-doi-org.brum.beds.ac.uk/10.3390/toxins11030177

AMA Style

Duracova M, Klimentova J, Myslivcova Fucikova A, Zidkova L, Sheshko V, Rehulkova H, Dresler J, Krocova Z. Targeted Mass Spectrometry Analysis of Clostridium perfringens Toxins. Toxins. 2019; 11(3):177. https://0-doi-org.brum.beds.ac.uk/10.3390/toxins11030177

Chicago/Turabian Style

Duracova, Miloslava, Jana Klimentova, Alena Myslivcova Fucikova, Lenka Zidkova, Valeria Sheshko, Helena Rehulkova, Jiri Dresler, and Zuzana Krocova. 2019. "Targeted Mass Spectrometry Analysis of Clostridium perfringens Toxins" Toxins 11, no. 3: 177. https://0-doi-org.brum.beds.ac.uk/10.3390/toxins11030177

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