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Isolation and analysis of peptidic fragments of alpha-gliadin using reversed-phase high-performance liquid chromatography
P Kocna, P Fric, M Kocova-Holakova, J Slaby, E Kasafirek, WT Hekkens
Jazyk angličtina Země Nizozemsko
- MeSH
- ektroforéza na škrobovém gelu MeSH
- gliadin * analýza MeSH
- peptidové fragmenty * analýza MeSH
- rostlinné proteiny * analýza MeSH
- vysokoúčinná kapalinová chromatografie MeSH
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.
Citace poskytuje Crossref.org
Literatura
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- $a Kocna, Petr, $d 1955- $7 jo2003181502 $u Laboratory of Gastroenterology, Charles University, Faculty of General Medicine, Prague, Czechoslovakia.
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- $a Isolation and analysis of peptidic fragments of alpha-gliadin using reversed-phase high-performance liquid chromatography / $c P Kocna, P Fric, M Kocova-Holakova, J Slaby, E Kasafirek, WT Hekkens
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- $a 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.
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