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A simple and inexpensive method for practical storage of field-sample proteins for subsequent MALDI-TOF MS analysis
BACKGROUND: Protein-containing samples can readily be characterised and/or identified using matrix-assisted laser-desorption and ionisation time-of-flight mass spectrometry (MALDI-TOF MS). This technique however requires relatively-fresh biological material that contains proteins that have not yet u...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6192001/ https://www.ncbi.nlm.nih.gov/pubmed/30356946 http://dx.doi.org/10.1186/s13007-018-0358-8 |
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author | Reeve, Michael A. Buddie, Alan G. |
author_facet | Reeve, Michael A. Buddie, Alan G. |
author_sort | Reeve, Michael A. |
collection | PubMed |
description | BACKGROUND: Protein-containing samples can readily be characterised and/or identified using matrix-assisted laser-desorption and ionisation time-of-flight mass spectrometry (MALDI-TOF MS). This technique however requires relatively-fresh biological material that contains proteins that have not yet undergone significant degradation. For field-work collection of samples, problems are often encountered due to delays between collection and sample processing, sample storage (possibly at elevated temperature and/or humidity in some climates), quarantine/regulatory restrictions on the transfer of living biological materials across national borders, and the potential to transfer unwanted microorganisms via non-living biological materials. RESULTS: In an attempt to overcome the above difficulties, we have developed a simple and inexpensive method for practical storage of field-sample proteins, for subsequent MALDI-TOF MS analysis, in which biological material is crushed onto filter paper and dried. The dried and protein-impregnated filter paper can then be soaked in an alcoholic solution suitable for the inactivation of microorganisms of concern and again dried for storage. After subsequent dry storage, the proteins may be eluted from the paper using a solution containing acetonitrile, trifluoroacetic acid, water, and MALDI-TOF MS matrix near to saturation. The extracted proteins are then pipetted onto the MALDI-TOF MS sample plate for subsequent analysis. Using this method, spectra of comparable quality to fresh-material controls have been obtained for acid-soluble proteins from Fallopia japonica and Impatiens glandulifera leaf material. Unlike untreated leaf material, high-quality spectra can be obtained with and without alcohol treatment even after storage for one month at up to 40 °C. CONCLUSIONS: We have developed a simple and inexpensive method for practical storage of field-sample proteins for subsequent MALDI-TOF MS analysis. Key benefits of this approach are a reduction in sample degradation, and consequent conservation of taxon-discriminatory spectral profiles, whilst minimising the potential for carryover of viable microorganisms. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13007-018-0358-8) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6192001 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-61920012018-10-23 A simple and inexpensive method for practical storage of field-sample proteins for subsequent MALDI-TOF MS analysis Reeve, Michael A. Buddie, Alan G. Plant Methods Methodology BACKGROUND: Protein-containing samples can readily be characterised and/or identified using matrix-assisted laser-desorption and ionisation time-of-flight mass spectrometry (MALDI-TOF MS). This technique however requires relatively-fresh biological material that contains proteins that have not yet undergone significant degradation. For field-work collection of samples, problems are often encountered due to delays between collection and sample processing, sample storage (possibly at elevated temperature and/or humidity in some climates), quarantine/regulatory restrictions on the transfer of living biological materials across national borders, and the potential to transfer unwanted microorganisms via non-living biological materials. RESULTS: In an attempt to overcome the above difficulties, we have developed a simple and inexpensive method for practical storage of field-sample proteins, for subsequent MALDI-TOF MS analysis, in which biological material is crushed onto filter paper and dried. The dried and protein-impregnated filter paper can then be soaked in an alcoholic solution suitable for the inactivation of microorganisms of concern and again dried for storage. After subsequent dry storage, the proteins may be eluted from the paper using a solution containing acetonitrile, trifluoroacetic acid, water, and MALDI-TOF MS matrix near to saturation. The extracted proteins are then pipetted onto the MALDI-TOF MS sample plate for subsequent analysis. Using this method, spectra of comparable quality to fresh-material controls have been obtained for acid-soluble proteins from Fallopia japonica and Impatiens glandulifera leaf material. Unlike untreated leaf material, high-quality spectra can be obtained with and without alcohol treatment even after storage for one month at up to 40 °C. CONCLUSIONS: We have developed a simple and inexpensive method for practical storage of field-sample proteins for subsequent MALDI-TOF MS analysis. Key benefits of this approach are a reduction in sample degradation, and consequent conservation of taxon-discriminatory spectral profiles, whilst minimising the potential for carryover of viable microorganisms. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13007-018-0358-8) contains supplementary material, which is available to authorized users. BioMed Central 2018-10-15 /pmc/articles/PMC6192001/ /pubmed/30356946 http://dx.doi.org/10.1186/s13007-018-0358-8 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Methodology Reeve, Michael A. Buddie, Alan G. A simple and inexpensive method for practical storage of field-sample proteins for subsequent MALDI-TOF MS analysis |
title | A simple and inexpensive method for practical storage of field-sample proteins for subsequent MALDI-TOF MS analysis |
title_full | A simple and inexpensive method for practical storage of field-sample proteins for subsequent MALDI-TOF MS analysis |
title_fullStr | A simple and inexpensive method for practical storage of field-sample proteins for subsequent MALDI-TOF MS analysis |
title_full_unstemmed | A simple and inexpensive method for practical storage of field-sample proteins for subsequent MALDI-TOF MS analysis |
title_short | A simple and inexpensive method for practical storage of field-sample proteins for subsequent MALDI-TOF MS analysis |
title_sort | simple and inexpensive method for practical storage of field-sample proteins for subsequent maldi-tof ms analysis |
topic | Methodology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6192001/ https://www.ncbi.nlm.nih.gov/pubmed/30356946 http://dx.doi.org/10.1186/s13007-018-0358-8 |
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