Purification of Parvalbumin from Carp

A Protocol that Avoids Heat-Treatment

Stef J. Koppelman, Roland A. Romijn, Harmen H.J. de Jongh, Julie A. Nordlee, Sander Piersma, Martin Hessing, Stephen L Taylor

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Parvalbumin from carp, a major allergen, was purified to homogeneity using ion exchange chromatography and size exclusion chromatography (estimated purity >95% to 98% based on SDS-PAGE and native PAGE) with a yield of 318 mg, and a number of basic biochemical characteristics were determined. The identity was confirmed by peptide-mass fingerprinting, and IgE-binding was demonstrated. The UV/Vis absorbance spectra were explained using the previously published amino acid sequences. Far UV-CD spectroscopy was used to confirm the folding character of parvalbumin. We conclude that parvalbumin from carp can be purified on a comparatively large (hundreds of milligrams) scale using a purification protocol that does not include denaturing steps. The purified protein resembles biochemical characteristics as were earlier published for carp parvalbumin, that is, a molecular weight of approximately 12 kDa, amino acid sequence identity and a secondary structure containing α-helices and β-structures. The described method provides a yield sufficient to produce and characterize antibodies to construct immunochemical methods to detect parvalbumin in food, as well as for use as a standard calibrator for such assays.Practical Application: Parvalbumin is a major allergen from fish. Here, we have purified a comparatively large quantity from carp that can be used to develop antisera for use in an assay method to detect fish allergens.

Original languageEnglish (US)
JournalJournal of food science
Volume75
Issue number3
DOIs
StatePublished - Apr 1 2010

Fingerprint

Parvalbumins
Carps
carp
Hot Temperature
allergens
heat treatment
Allergens
amino acid sequences
circular dichroism spectroscopy
Amino Acid Sequence
Fishes
assays
ion exchange chromatography
fish
purity
absorbance
antiserum
Native Polyacrylamide Gel Electrophoresis
Peptide Mapping
methodology

Keywords

  • Allergen
  • Carp
  • Fish
  • Parvalbumin

ASJC Scopus subject areas

  • Food Science

Cite this

Koppelman, S. J., Romijn, R. A., de Jongh, H. H. J., Nordlee, J. A., Piersma, S., Hessing, M., & Taylor, S. L. (2010). Purification of Parvalbumin from Carp: A Protocol that Avoids Heat-Treatment. Journal of food science, 75(3). https://doi.org/10.1111/j.1750-3841.2010.01513.x

Purification of Parvalbumin from Carp : A Protocol that Avoids Heat-Treatment. / Koppelman, Stef J.; Romijn, Roland A.; de Jongh, Harmen H.J.; Nordlee, Julie A.; Piersma, Sander; Hessing, Martin; Taylor, Stephen L.

In: Journal of food science, Vol. 75, No. 3, 01.04.2010.

Research output: Contribution to journalArticle

Koppelman SJ, Romijn RA, de Jongh HHJ, Nordlee JA, Piersma S, Hessing M et al. Purification of Parvalbumin from Carp: A Protocol that Avoids Heat-Treatment. Journal of food science. 2010 Apr 1;75(3). https://doi.org/10.1111/j.1750-3841.2010.01513.x
Koppelman, Stef J. ; Romijn, Roland A. ; de Jongh, Harmen H.J. ; Nordlee, Julie A. ; Piersma, Sander ; Hessing, Martin ; Taylor, Stephen L. / Purification of Parvalbumin from Carp : A Protocol that Avoids Heat-Treatment. In: Journal of food science. 2010 ; Vol. 75, No. 3.
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