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An antibody-free sample pretreatment method for osteopontin combined with MALDI-TOF MS/MS analysis

Osteopontin is an osteoblast-secreted protein with an aspartic acid-rich, highly phosphorylated, and glycosylated structure. Osteopontin can easily bind to integrins, tumor cells, extracellular matrix and calcium, and is related to bone diseases, various cancers, inflammation etc. Here, DEAE-Cibacro...

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Detalles Bibliográficos
Autores principales: Zhou, Yuye, Romson, Joakim, Emmer, Åsa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6405093/
https://www.ncbi.nlm.nih.gov/pubmed/30845167
http://dx.doi.org/10.1371/journal.pone.0213405
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author Zhou, Yuye
Romson, Joakim
Emmer, Åsa
author_facet Zhou, Yuye
Romson, Joakim
Emmer, Åsa
author_sort Zhou, Yuye
collection PubMed
description Osteopontin is an osteoblast-secreted protein with an aspartic acid-rich, highly phosphorylated, and glycosylated structure. Osteopontin can easily bind to integrins, tumor cells, extracellular matrix and calcium, and is related to bone diseases, various cancers, inflammation etc. Here, DEAE-Cibacron blue 3GA was used to extract recombinant osteopontin from human plasma, and to deplete abundant plasma proteins with an antibody-free method. Using selected buffer systems, osteopontin and human serum albumin could be bound to DEAE-Cibacron blue 3GA, while immunoglobulin G was excluded. The bound osteopontin could then be separated from albumin by using different sequential elution buffers. By this method, 1 μg/mL recombinant osteopontin could be separated from the major part of the most abundant proteins in human plasma. After trypsin digestion, the extracted osteopontin could be successfully detected and identified by MALDI-TOF MS/MS using the m/z 1854.898 peptide and its fragments.
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spelling pubmed-64050932019-03-17 An antibody-free sample pretreatment method for osteopontin combined with MALDI-TOF MS/MS analysis Zhou, Yuye Romson, Joakim Emmer, Åsa PLoS One Research Article Osteopontin is an osteoblast-secreted protein with an aspartic acid-rich, highly phosphorylated, and glycosylated structure. Osteopontin can easily bind to integrins, tumor cells, extracellular matrix and calcium, and is related to bone diseases, various cancers, inflammation etc. Here, DEAE-Cibacron blue 3GA was used to extract recombinant osteopontin from human plasma, and to deplete abundant plasma proteins with an antibody-free method. Using selected buffer systems, osteopontin and human serum albumin could be bound to DEAE-Cibacron blue 3GA, while immunoglobulin G was excluded. The bound osteopontin could then be separated from albumin by using different sequential elution buffers. By this method, 1 μg/mL recombinant osteopontin could be separated from the major part of the most abundant proteins in human plasma. After trypsin digestion, the extracted osteopontin could be successfully detected and identified by MALDI-TOF MS/MS using the m/z 1854.898 peptide and its fragments. Public Library of Science 2019-03-07 /pmc/articles/PMC6405093/ /pubmed/30845167 http://dx.doi.org/10.1371/journal.pone.0213405 Text en © 2019 Zhou et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Zhou, Yuye
Romson, Joakim
Emmer, Åsa
An antibody-free sample pretreatment method for osteopontin combined with MALDI-TOF MS/MS analysis
title An antibody-free sample pretreatment method for osteopontin combined with MALDI-TOF MS/MS analysis
title_full An antibody-free sample pretreatment method for osteopontin combined with MALDI-TOF MS/MS analysis
title_fullStr An antibody-free sample pretreatment method for osteopontin combined with MALDI-TOF MS/MS analysis
title_full_unstemmed An antibody-free sample pretreatment method for osteopontin combined with MALDI-TOF MS/MS analysis
title_short An antibody-free sample pretreatment method for osteopontin combined with MALDI-TOF MS/MS analysis
title_sort antibody-free sample pretreatment method for osteopontin combined with maldi-tof ms/ms analysis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6405093/
https://www.ncbi.nlm.nih.gov/pubmed/30845167
http://dx.doi.org/10.1371/journal.pone.0213405
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