Purification of selenoprotein P from human plasma

Biochim Biophys Acta. 1994 Feb 16;1204(2):243-9. doi: 10.1016/0167-4838(94)90014-0.


Selenoprotein P was partially purified (> 1000-fold) from human plasma in four chromatographic steps using 75Se-labeled selenoprotein P secreted by HepG2 cells in culture as a marker. The purified preparation was injected into mice and monoclonal antibodies, which precipitated the labeled protein, were generated. Neither of two different monoclonal antibodies had cross-reactivity with plasma from five animal species. Antibodies were coupled to agarose, and selenoprotein P was purified from human plasma by immunoaffinity chromatography followed by chromatography on heparin agarose. With two different matrix-bound monoclonal antibodies, the purification procedure gave two bands on SDS-PAGE with mobilities corresponding to 61 and 55 kDa. Both bands stained for carbohydrate and showed increased electrophoretic mobility after enzymatic deglycosylation. Immunoaffinity chromatography removed approx. one-third of the selenium from plasma or 0.4 mumol Se/l at a total selenium concentration of 1.1 mumol/l, indicating that selenoprotein P constituted this proportion of total plasma selenium in healthy US blood donors.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Monoclonal
  • Cell Line
  • Chromatography, Affinity
  • Hepatoblastoma / chemistry
  • Humans
  • Molecular Sequence Data
  • Proteins / chemistry
  • Proteins / immunology
  • Proteins / isolation & purification*
  • Selenium / blood
  • Selenium / isolation & purification
  • Selenoprotein P
  • Selenoproteins


  • Antibodies, Monoclonal
  • Proteins
  • Selenoprotein P
  • Selenoproteins
  • Selenium