Separation of human apolipoproteins A-IV, A-I and E by reversed-phase high-performance liquid chromatography on a TSK Phenyl-5PW column.
A method has been developed for the rapid separation of the medium-molecular-weight apolipoproteins A-IV, A-I and E by high-performance liquid chromatography. Separations were achieved using a commercially available column of very low hydrophobicity (TSK Phenyl-5PW) in the reversed-phase mode rather than the conventional mode of hydrophobic interaction. Delipidated apolipoproteins were dissolved in 20 mM orthophosphoric acid (pH 2.3), applied to the column which was pre-equilibrated with the same buffer, and eluted with an increasing gradient of acetonitrile. Purified apolipoproteins were identified by a combination of sodium dodecyl sulphate-polyacrylamide gel electrophoresis, amino acid analysis and N-terminal sequence analysis. In one step the method can be used to separate the major human chylomicron apolipoproteins A-IV, A-I and E, following preliminary removal of apolipoprotein A-II and the C apolipoproteins by size-exclusion chromatography.
Tetaz
T
T
Baker Medical Research Institute, Melbourne, Victoria, Australia.
Kecorius
E
E
Grego
B
B
Fidge
N
N
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D016632:Apolipoprotein A-I; D001054:Apolipoproteins A / Q000302:isolation & purification*; D001057:Apolipoproteins E / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D002914:Chylomicrons / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007202:Indicators and Reagents; D008196:Lymph / Q000737:chemistry; D008969:Molecular Sequence Data
D000596:Amino Acids; D016632:Apolipoprotein A-I; D001054:Apolipoproteins A; D001057:Apolipoproteins E; D002914:Chylomicrons; D007202:Indicators and Reagents; C022205:apolipoprotein A-IV