Purification and analysis of a 5 kDa component of enamel matrix derivative

Alexandra Mumulidu, Bergisa Hildebrand, Beata Fabi, Lars Hammarström, David L. Cochran, Michel Dard, Stephanie Lemoult

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

High performance liquid chromatography (HPLC) methods were used to analyse a 5 kDa component purified from enamel matrix derivative (EMD), the active ingredient in Emdogain®, a commercial product for periodontal tissue regeneration. After initial purification by size-exclusion chromatography (SEC) on a 100 cm × 5 cm column (Bio-Gel P-30 Fine, 280 nm), collected fractions were analysed by size-exclusion HPLC (SE HPLC; TSK-Gel Super SW2000, 220 nm). The fractions containing only the 5 kDa component were analysed by reversed-phase high-pressure chromatography (RP HPLC; YMC-Pack ODS-A, 200 nm), revealing four peaks of the 5 kDa component. From 1200 mg of EMD (of which 9% is the 5 kDa component), approximately 65 mg of lyophilised 5 kDa component were obtained, corresponding to a recovery of 60%. The SE HPLC method was mainly suitable for qualitative analysis, whereas the RP HPLC method was appropriate for both qualitative and quantitative analysis.

Original languageEnglish (US)
Pages (from-to)210-218
Number of pages9
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume857
Issue number2
DOIs
StatePublished - Oct 1 2007

Keywords

  • Emdogain
  • Enamel matrix derivative
  • Fractionation
  • Method validation
  • Protein purification
  • Qualitative analysis
  • Quantitative analysis
  • Reversed-phase HPLC
  • Size-exclusion HPLC

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology

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