Expanded bed adsorption as a fast technique for the large-scale purification of the complete isoform pool of Ber e 1, the major allergen from Brazil nuts
article
A new, fast, large-scale purification method for Ber e 1, the major allergen from Brazil nuts, using expanded bed adsorption (EBA) chromatography, is presented. Using EBA, crude extracts can be applied to a fluidized column, which allows the unhindered passage of particulate impurities, thereby avoiding time-consuming centrifugation or filtration steps. With this new purification method, 2.8 g of Ber e 1 was obtained from 85 g defatted Brazil nut meal, essentially within 1 day. Various structural as well as immunochemical characteristics of the purified protein were determined, and compared to those of Ber e 1 purified using conventional chromatographic techniques. The complete pool of Ber e 1 isoforms was collected using EBA. The most abundant isoforms were observed to have pI around 8 and heterogeneity was observed in both the large and the small subunit of the heterodimeric protein. Ber e 1 has a highly ordered secondary structure. No apparent differences in immune reactivity were observed between EBA purified Ber e 1 and conventionally purified Ber e 1, using IgE-binding experiments. Thus, using EBA, Bere 1 can be purified fast and on gram-scale, while having purity equal to that of conventionally purified Ber e 1. © 2006 WILEY-VCH Verlag GmbH & Co. KGaA.
Topics
BiotechnologyAllergenBer e 1ChromatographyExpanded bed adsorptionPurification2S albumin precursor, Bertholletia excelsaAlbuminoidAllergenProtein precursorVegetable proteinAdsorptionBrazil nutChemistryChromatographyCircular dichroismIsolation and purificationMass spectrometryMethodologyPlant seedPolyacrylamide gel electrophoresisAdsorptionAlbuminsAllergensBertholletiaChromatographyCircular DichroismElectrophoresis, Polyacrylamide GelPlant ProteinsProtein PrecursorsSeedsSpectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationBertholletia excelsaHuman echovirus 1Ziziphus mauritiana
TNO Identifier
239143
ISSN
16134125
Source
Molecular Nutrition and Food Research, 50(3), pp. 275-281.
Pages
275-281
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