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

Waga, J.; Zientarski, J.,Plant Breeding and Acclimatization Institute, Radzikow . Dept. of Cereals Breeding and Quality Evaluation, Krakow

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

2007

Waga, J. | Zientarski, J.,Plant Breeding and Acclimatization Institute, Radzikow (Poland). Dept. of Cereals Breeding and Quality Evaluation, Krakow

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 alpha and beta gliadins, while the immunoenzymatic reaction with gamma and omega-gliadins was much weaker

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