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An antibody-free sample pretreatment method for osteopontin combined with MALDI-TOF MS/MS analysis
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemistry, Applied Physical Chemistry.ORCID iD: 0000-0002-0186-7795
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemistry, Applied Physical Chemistry.ORCID iD: 0000-0003-3091-3882
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemistry, Applied Physical Chemistry.ORCID iD: 0000-0002-3444-9987
2019 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 14, no 3, article id e0213405Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
Public Library of Science , 2019. Vol. 14, no 3, article id e0213405
National Category
Analytical Chemistry
Research subject
Chemistry
Identifiers
URN: urn:nbn:se:kth:diva-249443DOI: 10.1371/journal.pone.0213405ISI: 000460638800082PubMedID: 30845167Scopus ID: 2-s2.0-85062607743OAI: oai:DiVA.org:kth-249443DiVA, id: diva2:1304376
Note

QC 20190522

Available from: 2019-04-12 Created: 2019-04-12 Last updated: 2019-05-22Bibliographically approved

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Publisher's full textPubMedScopushttps://doi.org/10.1371/journal.pone.0213405

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Zhou, YuyeRomson, JoakimEmmer, Åsa

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