AI Article Synopsis
- Osteopontin is a protein secreted by osteoblasts that plays a role in bone diseases, cancers, and inflammation due to its ability to bind various molecules.
- DEAE-Cibacron blue 3GA was utilized to extract recombinant osteopontin from human plasma, effectively removing major abundant proteins without using antibodies.
- The process involved using specific buffer systems to separate osteopontin from other proteins, which was later identified using MALDI-TOF MS/MS after digestion with trypsin.
Article Abstract
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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Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6405093 | PMC |
| http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0213405 | PLOS |
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