title: Preparative peptide purification by cation-exchange and reversed-phase perfusion chromatography.
abstract: 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.
authors.affiliation: PerSeptive Biosystems, Cambridge, MA 02139.
authors.forename: S
authors.initials: S
authors.lastname: Fulton
authors.forename: M
authors.initials: M
authors.lastname: Meys
authors.forename: J
authors.initials: J
authors.lastname: Protentis
authors.forename: N B
authors.initials: NB
authors.lastname: Afeyan
authors.forename: J
authors.initials: J
authors.lastname: Carlton
authors.forename: J
authors.initials: J
authors.lastname: Haycock
mesh_terms: 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*
chemical_list: D014660:Vasoactive Intestinal Peptide