title: Isolation and analysis of peptidic fragments of alpha-gliadin using reversed-phase high-performance liquid chromatography.
abstract: Peptidic fragments of alpha-gliadin were obtained by peptic-tryptic-pancreatic (PTP) digestion of the alpha-gliadin fraction isolated by ion-exchange chromatography on a sulphopropyl-Sephadex C-50 column. The proteolytic digest was fractionated by ultrafiltration into three subfractions, PTPa1-PTPa3. The subfraction PTPa2 was then analysed and individual peaks were separated using reversed-phase high-performance liquid chromatography (RP-HPLC) using a gradient of acetonitrile in 0.1% trifluoroacetic acid and a Separon SGX-C18 sorbent. A 100-mg amount of the PTPa2 subfraction was separated in a single analysis by preparative RP-HPLC and twenty peaks were obtained for further characterization. The molecular mass in range 300-3000 was established for individual peptidic fragments by gel-permeation chromatography on a TSK-G2000 SW column.
authors.affiliation: Laboratory of Gastroenterology, Charles University, Faculty of General Medicine, Prague, Czechoslovakia.
authors.forename: P
authors.initials: P
authors.lastname: Kocna
authors.forename: P
authors.initials: P
authors.lastname: Fric
authors.forename: M
authors.initials: M
authors.lastname: Kocová-Holáková
authors.forename: J
authors.initials: J
authors.lastname: Slabý
authors.forename: E
authors.initials: E
authors.lastname: Kasafírek
authors.forename: W T
authors.initials: WT
authors.lastname: Hekkens
mesh_terms: D002851:Chromatography, High Pressure Liquid; D004592:Electrophoresis, Starch Gel; D005903:Gliadin / Q000032:analysis*; D010446:Peptide Fragments / Q000032:analysis*; D010940:Plant Proteins / Q000032:analysis*
chemical_list: D010446:Peptide Fragments; D010940:Plant Proteins; D005903:Gliadin