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.

E.L. van Boxtel, G.A. van Koningsveld, S.J. Koppelman, L.A.M. van den Broek, A.G.J. Voragen, H. Gruppen

Research output: Contribution to journalArticleAcademicpeer-review

1 Citation (Scopus)

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, Ber e 1 can be purified fast and on gram-scale, while having purity equal to that of conventionally purified Ber e 1.
Original languageEnglish
Pages (from-to)275-281
JournalMolecular Nutrition & Food Research
Volume50
Issue number3
DOIs
Publication statusPublished - 2006

Fingerprint

Bertholletia
Brazil nuts
allergens
Allergens
Adsorption
Protein Isoforms
adsorption
purification methods
chromatography
methodology
Protein Subunits
protein subunits
Complex Mixtures
Centrifugation
purity
Immunoglobulin E
centrifugation
Meals
Chromatography
particulates

Keywords

  • bertholletia-excelsa hbk
  • sulfur-rich protein
  • 2s albumin
  • expression
  • peanut

Cite this

@article{e23700cc2e15446881a67fbcea2de970,
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.",
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, Ber e 1 can be purified fast and on gram-scale, while having purity equal to that of conventionally purified Ber e 1.",
keywords = "bertholletia-excelsa hbk, sulfur-rich protein, 2s albumin, expression, peanut",
author = "{van Boxtel}, E.L. and {van Koningsveld}, G.A. and S.J. Koppelman and {van den Broek}, L.A.M. and A.G.J. Voragen and H. Gruppen",
year = "2006",
doi = "10.1002/mnfr.200500203",
language = "English",
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pages = "275--281",
journal = "Molecular Nutrition & Food Research",
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}

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. / van Boxtel, E.L.; van Koningsveld, G.A.; Koppelman, S.J.; van den Broek, L.A.M.; Voragen, A.G.J.; Gruppen, H.

In: Molecular Nutrition & Food Research, Vol. 50, No. 3, 2006, p. 275-281.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - 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.

AU - van Boxtel, E.L.

AU - van Koningsveld, G.A.

AU - Koppelman, S.J.

AU - van den Broek, L.A.M.

AU - Voragen, A.G.J.

AU - Gruppen, H.

PY - 2006

Y1 - 2006

N2 - 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, Ber e 1 can be purified fast and on gram-scale, while having purity equal to that of conventionally purified Ber e 1.

AB - 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, Ber e 1 can be purified fast and on gram-scale, while having purity equal to that of conventionally purified Ber e 1.

KW - bertholletia-excelsa hbk

KW - sulfur-rich protein

KW - 2s albumin

KW - expression

KW - peanut

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