Title
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
Author
van Boxtel, E.L.
van Koningsveld, G.A.
Koppelman, S.J.
van den Broek, L.A.M.
Voragen, A.G.J.
Gruppen, H.
TNO Kwaliteit van Leven
Publication year
2006
Abstract
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.
Subject
Biology
Biotechnology
Allergen
Ber e 1
Chromatography
Expanded bed adsorption
Purification
2S albumin precursor, Bertholletia excelsa
albuminoid
allergen
protein precursor
vegetable protein
adsorption
article
Brazil nut
chemistry
chromatography
circular dichroism
isolation and purification
mass spectrometry
methodology
plant seed
polyacrylamide gel electrophoresis
Adsorption
Albumins
Allergens
Bertholletia
Chromatography
Circular Dichroism
Electrophoresis, Polyacrylamide Gel
Plant Proteins
Protein Precursors
Seeds
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Bertholletia excelsa
Human echovirus 1
Ziziphus mauritiana
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DOI
https://doi.org/10.1002/mnfr.200500203
TNO identifier
239143
ISSN
1613-4125
Source
Molecular Nutrition and Food Research, 50 (3), 275-281
Document type
article