Isolation and purification of individual gliadin proteins by preparative acid polyacrylamide gel electrophoresis [A-PAGE] for allergenic research

• Autorzy: Waga J. , Zientarski J.

Warianty tytułu

PL

Izolowanie i oczyszczanie alergizujacych bialek gliadynowych metoda elektroforezy preparatywnej [A-PAGE]

• Języki publikacji: EN

Abstrakty

EN

Gliadins are a large and complex group of proteins, consisting of dozens to hundreds of unique polypeptides in any wheat cultivar. Individual gliadins differ in molecular weight, isoelectric point, and amino acid sequences, making their analyses especially difficult. We here used preparative polyacrylamide gel electrophoresis in acidic conditions to fractionate gliadins of the cultivar Fraza. Resulting fractions were then analysed by ELISA to test their immunoreactive properties with antigliadin polyclonal antibodies. Preparative electrophoresis yielded 60 aliquots, of which 15 contained single proteins. Separated gliadins differed in immunoreactive properties: antigliadin antibodies bound stronger to a and b gliadins, while the immunoenzymatic reaction with g and w-gliadins was much weaker.

PL

Gliadyny stanowią rozbudowany kompleks białkowy, który – zależnie od odmiany pszenicy – tworzy od kilkudziesięciu do kilkuset alergizujących monomerów. Poszczególne monomery różnią się masą cząsteczkową, punktem izoelektrycznym, a przede wszystkim sekwencjami aminokwasów, co znacznie utrudnienia analizę właściwości alergogennych. W niniejszej pracy wykorzystano zmodyfikowaną metodę elektroforezy preparatywnej na żelu poliakryloamidowym w środowisku kwaśnym do rozdziału gliadyn pszenicy ozimej Fraza. Oczyszczone białka analizowano następnie pod względem właściwości immunoreaktywnych testem ELISA w reakcji z poliklonalnymi przeciwciałami antygliadynowymi. Uzyskano 60 frakcji, z których 15 zawierało pojedyncze monomery. Stwierdzono zróżnicowanie immunoreaktywności rozdzielonych białek – przeciwciała antygliadynowe silnie wiązały gliadyny a i b natomiast reaktywność g i w gliadyn była znacznie niższa.

Słowa kluczowe

EN

acid polyacrylamide gel electrophoresis; wheat; gliadin protein; isolation; ELISA test; purification; protein; immunoreactivity; allergenicity

• Wydawca: -
• Czasopismo: Polish Journal of Food and Nutrition Sciences
• Rocznik: 2007
• Tom: 57
• Numer: 1
• Opis fizyczny: p.91-95,fig.,ref.

Twórcy

autor

Waga J.

• Plant Breeding and Acclimatisation Institute, Zawila 4, 30-423 Krakow, Poland

autor

Zientarski J.

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