{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,17]],"date-time":"2026-06-17T01:51:56Z","timestamp":1781661116192,"version":"3.54.5"},"reference-count":18,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2010,11,22]],"date-time":"2010-11-22T00:00:00Z","timestamp":1290384000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Proteomics Clinical Apps"],"published-print":{"date-parts":[[2010,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p><jats:bold>Purpose:<\/jats:bold> Hemoglobin (Hb) depletion with nickel affinity chromatography has been shown to increase the number of proteins identified in proteomic studies of erythrocytes, but limited data exist on the application of this technique in depletion of Hb from plasma or serum required for clinical biomarker studies. The aim of this study was to explore the potential of using nickel\u2010beads for Hb depletion of plasma.<\/jats:p><jats:p><jats:bold>Experimental design:<\/jats:bold> Nickel\u2013nitrilotriacetic acid (Ni\u2013NTA) affinity chromatography was used to deplete Hb from hemolyzed plasma samples obtained from children with sickle cell disease (SCD, <jats:italic>n<\/jats:italic>=7) and normal human plasma (<jats:italic>n<\/jats:italic>=4). Ni\u2013NTA\u2010bound proteins were analyzed by one\u2010dimensional GE, followed by in\u2010gel digestion for characterization using an LTQ\u2010Orbitrap hybrid mass spectrometer. In addition, the loss of two non\u2010Hb\u2010related plasma proteins, thrombospondin1 and <jats:sc>L<\/jats:sc>\u2010selectin, by Ni\u2013NTA was determined by ELISA (SCD <jats:italic>n<\/jats:italic>=6, non\u2010SCD controls <jats:italic>n<\/jats:italic>=2).<\/jats:p><jats:p><jats:bold>Results:<\/jats:bold> Ni\u2013NTA resulted in an average 60% decrease in plasma protein concentration, which was not hemolysis dependent. Specifically, Hb (7 peptides) and the top three proteins, \u03b1\u20102\u2010macroglobulin (75 peptides), apolipoprotein B\u2010100 (73 peptides), and albumin (42 peptides) were Ni\u2013NTA bound. In addition, using an ELISA assay two non\u2010Hb\u2010associated plasma proteins thrombospondin1 and <jats:sc>L<\/jats:sc>\u2010selectin were decreased by Ni\u2010NTA.<\/jats:p><jats:p><jats:bold>Conclusions and clinical relevance:<\/jats:bold> Hb depletion with Ni\u2013NTA is effective for Hb removal but is not specific. There is a potential for deleterious depletion of potential biomarkers that may limit the applicability of this method. Consideration of alternate methods of Hb depletion for clinical proteomics may be warranted.<\/jats:p>","DOI":"10.1002\/prca.201000054","type":"journal-article","created":{"date-parts":[[2010,10,5]],"date-time":"2010-10-05T14:12:18Z","timestamp":1286287938000},"page":"926-930","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Hemoglobin depletion from plasma: Considerations for proteomic discovery in Sickle Cell disease and other hemolytic processes"],"prefix":"10.1002","volume":"4","author":[{"given":"Lisa M.","family":"Williams","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zongming","family":"Fu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pratima","family":"Dulloor","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy","family":"Yen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emily","family":"Barron\u2010Casella","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William","family":"Savage","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer E.","family":"Van Eyk","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James 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