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- Title
Fractionation of selenium-containing proteins in serum by multiaffinity liquid chromatography before size-exclusion chromatography–ICPMS.
- Authors
Palacios, Òscar; Encinar, Jorge Ruiz; Schaumlöffel, Dirk; Lobinski, Ryszard
- Abstract
Immunoaffinity chromatography has been investigated for fractionation of serum into selenoalbumin and true selenoproteins. Among several albumin-depletion kits tested, a multiaffinity column specifically binding albumin and five other major serum proteins provided the best results. It extracted ca 95% of both albumin and selenoalbumin, which enabled interference-free determination of glutathione peroxidase, selenoprotein P, and selenoalbumin by size-exclusion chromatography combined with inductively coupled plasma mass spectrometry (SEC–ICPMS). The efficiency of the multiaffinity column did not vary over a period of 18 months. The purity of fractions separated by immunoaffinity LC was confirmed by elution-volume matching with standards in SEC–ICPMS and by selenopeptide mapping in capillary HPLC–ICPMS. Quantification of the selenium distribution among the different proteins in human serum from a control group and from a person on a selenium-rich diet revealed that 67% of the supplemented selenium was incorporated into albumin, 30% into glutathione peroxidase, and 3% into selenoprotein P.
- Subjects
LIQUID chromatography; BLOOD protein separation; SELENOPROTEINS; ALBUMINS; GLUTATHIONE
- Publication
Analytical & Bioanalytical Chemistry, 2006, Vol 384, Issue 6, p1276
- ISSN
1618-2642
- Publication type
Article
- DOI
10.1007/s00216-005-0286-0