BOČÁNEK, Ondřej, Ondrej ŠEDO, Stanislav PEKÁR and Zbyněk ZDRÁHAL. Evaluation of sample preparation protocols for spider venom profiling by MALDI-TOF MS. Toxicon. Oxford, England: PERGAMON-ELSEVIER SCIENCE LTD, 2017, vol. 133, July, p. 18-25. ISSN 0041-0101. Available from: https://dx.doi.org/10.1016/j.toxicon.2017.04.008.
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Basic information
Original name Evaluation of sample preparation protocols for spider venom profiling by MALDI-TOF MS
Authors BOČÁNEK, Ondřej (203 Czech Republic, belonging to the institution), Ondrej ŠEDO (203 Czech Republic, belonging to the institution), Stanislav PEKÁR (703 Slovakia, guarantor, belonging to the institution) and Zbyněk ZDRÁHAL (203 Czech Republic, belonging to the institution).
Edition Toxicon, Oxford, England, PERGAMON-ELSEVIER SCIENCE LTD, 2017, 0041-0101.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10406 Analytical chemistry
Country of publisher United Kingdom of Great Britain and Northern Ireland
Confidentiality degree is not subject to a state or trade secret
Impact factor Impact factor: 2.352
RIV identification code RIV/00216224:14310/17:00094825
Organization unit Faculty of Science
Doi http://dx.doi.org/10.1016/j.toxicon.2017.04.008
UT WoS 000403864000003
Keywords in English Venom mass fingerprinting; Spider venom; MALDI-TOF MS
Tags CF PROT, NZ, rivok
Changed by Changed by: Ing. Nicole Zrilić, učo 240776. Changed: 3/4/2018 14:56.
Abstract
Spider venoms are highly complex mixtures containing biologically active substances with potential for use in biotechnology or pharmacology. Fingerprinting of venoms by Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) is a thriving technology, enabling the rapid detection of peptide/protein components that can provide comparative information. In this study, we evaluated the effects of sample preparation procedures on MALDI-TOF mass spectral quality to establish a protocol providing the most reliable analytical outputs. We adopted initial sample preparation conditions from studies already published in this field. Three different MALDI matrixes, three matrix solvents, two sample deposition methods, and different acid concentrations were tested. As a model sample, venom from Brachypelma albopilosa was used. The mass spectra were evaluated on the basis of absolute and relative signal intensities, and signal resolution. By conducting three series of analyses at three weekly intervals, the reproducibility of the mass spectra were assessed as a crucial factor in the selection for optimum conditions. A sample preparation protocol based on the use of an HCCA matrix dissolved in 50% acetonitrile with 2.5% TFA deposited onto the target by the dried-droplet method was found to provide the best results in terms of information yield and repeatability. We propose that this protocol should be followed as a standard procedure, enabling the comparative assessment of MALDI-TOF MS spider venom fingerprints.
Links
GA15-14762S, research and development projectName: Koevoluce potravní-specializace a složení jedu u pavouků
Investor: Czech Science Foundation
LM2015043, research and development projectName: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)
Investor: Ministry of Education, Youth and Sports of the CR
LQ1601, research and development projectName: CEITEC 2020 (Acronym: CEITEC2020)
Investor: Ministry of Education, Youth and Sports of the CR
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