Title of article :
Preparative reversed-phase high-performance liquid chromatography collection efficiency for an antimicrobial peptide on columns of varying diameters (1 mm to 9.4 mm I.D.)
Author/Authors :
Chen، نويسنده , , Yuxin and Mant، نويسنده , , Colin T. and Hodges، نويسنده , , Robert S.، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2007
Abstract :
The present study examines the effect of reversed-phase high-performance liquid chromatography (RP-HPLC) column diameter (1 mm to 9.4 mm I.D.) on the one-step slow gradient preparative purification of a 26-residue synthetic antimicrobial peptide. When taken together, the semi-preparative column (9.4 mm I.D.) provided the highest yields of purified product (an average of 90.7% recovery from hydrophilic and hydrophobic impurities) over a wide range of sample load (0.75–200 mg). Columns with smaller diameters, such as narrowbore columns (150 × 2.1 mm I.D.) and microbore columns (150 × 1.0 mm I.D.), can be employed to purify peptides with reasonable recovery of purified product but the range of the crude peptide that can be applied to the column is limited. In addition, the smaller diameter columns require more extensive fraction analysis to locate the fractions of pure product than the larger diameter column with the same load. Our results show the excellent potential of the one-step slow gradient preparative protocol as a universal method for purification of synthetic peptides.
Keywords :
Reversed-phase high-performance liquid chromatography , antimicrobial peptide , Preparative chromatography
Journal title :
Journal of Chromatography A
Journal title :
Journal of Chromatography A