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 language | English (US) |
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Pages (from-to) | 210-218 |
Number of pages | 9 |
Journal | Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences |
Volume | 857 |
Issue number | 2 |
DOIs | |
State | Published - 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