TY - JOUR
T1 - Purification and analysis of a 5 kDa component of enamel matrix derivative
AU - Mumulidu, Alexandra
AU - Hildebrand, Bergisa
AU - Fabi, Beata
AU - Hammarström, Lars
AU - Cochran, David L.
AU - Dard, Michel
AU - Lemoult, Stephanie
PY - 2007/10/1
Y1 - 2007/10/1
N2 - 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.
AB - 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.
KW - Emdogain
KW - Enamel matrix derivative
KW - Fractionation
KW - Method validation
KW - Protein purification
KW - Qualitative analysis
KW - Quantitative analysis
KW - Reversed-phase HPLC
KW - Size-exclusion HPLC
UR - http://www.scopus.com/inward/record.url?scp=34548834437&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34548834437&partnerID=8YFLogxK
U2 - 10.1016/j.jchromb.2007.07.017
DO - 10.1016/j.jchromb.2007.07.017
M3 - Article
C2 - 17689154
AN - SCOPUS:34548834437
SN - 1570-0232
VL - 857
SP - 210
EP - 218
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
IS - 2
ER -