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Extraction,isolation, and concentration of calprotectin antigen (S100A8/S100A9) fromgranulocytes.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Clinical Chemistry.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Clinical Chemistry.
2018 (English)In: Health Science Reports, Vol. 1, p. e35-, article id 10.1002/hsr2.35Article in journal (Refereed) Published
Abstract [en]

Background:Calprotectin is a promising biomarker for granulocyte activation. It is mainlymeasured in faeces as a marker for inflammatory bowel disease. A limitation is that there is nowidely accepted calibrator.Aim:To establish a method for purification of calprotectin from human granulocytes that iseasily reproducible, reliable, and could contribute to a better agreement between differentcalprotectin methods.Methods and results:Calprotectin was purified from granulocyte extracts using ion‐exchange chromatography. The granulocytes were separated from blood bags. The purity wasanalysed by analysing pixel density of a picture of the sodium dodecyl sulfate polyacrylamidegel electrophoresis and by size exclusion chromatography. The calprotectin concentration ofthe pure antigen solution was determined using Biuret method. The purity was >95% for 3 prep-arations, and their concentrations were 1079, 1080, and 1813 mg/L.Conclusion:It is possible to reproducibly prepare highly purified calprotectin antigen fromhuman granulocytes. The preparations can be used for preparing calibrators, controls forimmunological calprotectin assays, and immunisation for raising antibodies against humancalprotectin in hens.

Place, publisher, year, edition, pages
Wiley-Blackwell Publishing Inc., 2018. Vol. 1, p. e35-, article id 10.1002/hsr2.35
National Category
Clinical Laboratory Medicine
Identifiers
URN: urn:nbn:se:uu:diva-355220DOI: 10.1002/hsr2.35OAI: oai:DiVA.org:uu-355220DiVA, id: diva2:1227929
Available from: 2018-06-27 Created: 2018-06-27 Last updated: 2018-06-27

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