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New approaches to chromatographic purification of bovine dopamine-beta-hydroxylase.

Authors
  • Varlamov, V P1
  • Lopatin, S A
  • Ilyina, A V
  • Bannikova, G E
  • Chlenov, M A
  • Vasiyarov, G G
  • Mitrochkin, A A
  • Morgunova EYu
  • Pierrot, M
  • Reglier, M
  • 1 Centre Bioengineering, Russian Academy of Sciences, Vavilova, Moscow.
Type
Published Article
Journal
Journal of Chromatography A
Publisher
Elsevier
Publication Date
Sep 08, 1995
Volume
711
Issue
1
Pages
113–118
Identifiers
PMID: 7496482
Source
Medline
License
Unknown

Abstract

The use of the traditional scheme for the isolation of bovine dopamine-beta-hydroxylase (bDBH) from bovine adrenal medulla resulted in active but not pure bDBH, containing about 50% of admixtures. Immobilized metal chelate affinity chromatography on agarose modified with iminodiacetic acid residues and charged with cobalt ions was applied in the final stage to obtain more than 90% pure and active bDBH. Final purification of bDBH using step elution with 0-0.5 M methyl-D-mannoside in buffer solution from concanavalin A-Sepharose was studied. The determination of bDBH in various samples was performed using size-exclusion chromatography.

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