Purification of vitamin D binding protein from human plasma using high performance liquid chromatography

Geoffrey A. Taylor, Herbert N. Mazhindu, John B C Findlay, Munro Peacock

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Vitamin D binding protein/group-specific component was purified from human plasma by chromatographic techniques utilising high performance liquid chromatography and by traditional low pressure Chromatographic techniques alone. Use of high performance liquid chromatography considerably reduced the time taken to prepare pure vitamin D binding protein and increased the yield to 16% compared with 2.8% using the traditional methods. The vitamin D binding protein prepared by high performance liquid chromatography was shown to be highly pure by amino acid sequence, SDS gel electrophoresis and by antibody production. The amino acid sequence was confirmed and extended. The affinity constants of the high pressure liquid chromatography purified vitamin D binding protein for 25 hydroxycholecalciferol (25 OHD3) and 1,25 dihydroxycholecalciferol 1,25(OH)2D3 were 1.9 × 107 mol/l and 2.6 × 106 mol/l, respectively.

Original languageEnglish (US)
Pages (from-to)31-41
Number of pages11
JournalClinica Chimica Acta
Volume155
Issue number1
DOIs
StatePublished - Feb 28 1986
Externally publishedYes

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Vitamin D-Binding Protein
Plasma (human)
High performance liquid chromatography
Purification
High Pressure Liquid Chromatography
Amino Acid Sequence
High pressure liquid chromatography
Calcifediol
Amino Acids
Calcitriol
Electrophoresis
Antibody Formation
Gels
Pressure
Antibodies

Keywords

  • 25 Hydroxycholecalciferol
  • High performance liquid chromatography
  • Plasma
  • Purification
  • Vitamin D binding protein

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

Cite this

Purification of vitamin D binding protein from human plasma using high performance liquid chromatography. / Taylor, Geoffrey A.; Mazhindu, Herbert N.; Findlay, John B C; Peacock, Munro.

In: Clinica Chimica Acta, Vol. 155, No. 1, 28.02.1986, p. 31-41.

Research output: Contribution to journalArticle

Taylor, Geoffrey A. ; Mazhindu, Herbert N. ; Findlay, John B C ; Peacock, Munro. / Purification of vitamin D binding protein from human plasma using high performance liquid chromatography. In: Clinica Chimica Acta. 1986 ; Vol. 155, No. 1. pp. 31-41.
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