Preparative peptide purification by cation-exchange and reversed-phase perfusion chromatography.
Cation exchange was compared to reversed-phase chromatography for the preparative purification of a 28-residue peptide (vasoactive intestinal polypeptide) on the 100-mg scale. Optimized high-speed, high-resolution methods were developed for both chromatographic modes on POROS Perfusion Chromatography flow-through particle chromatography columns. While both methods appeared to provide similar purity, the cation exchange column had approximately ten times the loading capacity per unit column volume as the reversed-phase column. Five-minute methods for desalting the cation exchange-purified peptide and analysis of fractions were developed using small reversed-phase columns. The cation-exchange method was scaled up to process 95 mg of crude peptide in a 12-min run.
Fulton
S
S
PerSeptive Biosystems, Cambridge, MA 02139.
Meys
M
M
Protentis
J
J
Afeyan
N B
NB
Carlton
J
J
Haycock
J
J
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange; D008969:Molecular Sequence Data; D014660:Vasoactive Intestinal Peptide / Q000302:isolation & purification*
D014660:Vasoactive Intestinal Peptide