Print page Print page
Switch language
The Capital Region of Denmark - a part of Copenhagen University Hospital

Purification and characterization of osteopontin from human milk

Research output: Contribution to journalJournal articleResearch

  1. Purification and characterization of recombinant full-length and protease domain of murine MMP-9 expressed in Drosophila S2 cells

    Research output: Contribution to journalJournal articleResearchpeer-review

  2. Expression, purification and characterization of the cancer-germline antigen GAGE12I: A candidate for cancer immunotherapy

    Research output: Contribution to journalJournal articleResearchpeer-review

View graph of relations
Osteopontin (OPN) is expressed in many organs and tissues and has different biological properties related to different molecular forms in respect to size and posttranslational modifications. However, a purification procedure for authentic intact OPN as well as fragments of OPN from an accessible biological source is missing. A four-step procedure was used to purify OPN from human milk, based on its crystal growth inhibitory activity, including anion exchange chromatography, the elimination of casein, hydroxyapatite chromatography, and negative affinity chromatography. Purified OPN was further separated into its different molecular forms by means of a two-step procedure, involving size exclusion chromatography and reverse phase chromatography. A rabbit polyclonal antibody was raised to purified intact OPN and high M(r) OPN components; the immunoreactivity of both forms was almost equal when investigated by enzyme immunoassay (EIA). The procedures facilitate the purification of intact OPN and OPN fragments for purposes of standardization, preparation of monospecific antibodies, and functional studies.
Translated title of the contributionPurification and characterization of osteopontin from human milk.
Original languageEnglish
JournalProtein Expression and Purification
Issue number2
Pages (from-to)238-245
Number of pages8
Publication statusPublished - 2003

ID: 32550921