Abstract

Parvalbumin (PV) is a major fish allergen that is characterized by high water solubility, low molecular weight (i.e., ~12,000), acidic isoelectric point, heat stability and high phenylalanine content (compared with tyrosine and tryptophan). We used a fast, simple and efficient method to purify PV from Cyprinus carpio (carp). Heating, two runs of trichloroacetic acid precipitation and gel filtration chromatography were integrated in this method. Results of sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blot showed that the purified protein was PV and the purity of carp PV obtained using this method was more than 90%. Moreover, the PV was characterized by MALDI-TOF-TOF MS and an unusually high 260 to 280 nm run absorbance ratio in the final purification stages. The rapid purification method used in this study resulted in better purification and identification of PV compared with other methods. Further studies on PV detection in freshwater fish can be performed based on the results of this experiment.

Keywords:

Allergen, Cyprinus carpio, identification, parvalbumin, purification,

References

1. Aas, K., 1966. Studies of hypersensitivity to fish. A clinical study. Int. Arch. Allergy Appl. Immunol., 29: 346-363.
   CrossRef    PMid:5949057    
2. Aas, K., 1969. Elsayed, S. Characterization of a major allergen (cod). J. Allergy, 44: 333-343.
   CrossRef    
3. Aas, K., 1987. Fish Allergy and the Codfish Allergen Model. In: Brostoff, J. and S.J. Challacombe (Eds.), Food Allergy and Intolerance; Bailli`ere-Tindall, London, pp: 356-366.
   
4. Bernhisel-Broadbent, J., S.M. Scanlon and H.A. Sampson, 1992. Fish hypersensitivity. I. In vitro and oral challenge results in fish-allergic patients. J. Allergy Clin. Immun., 89: 730-737.
   CrossRef    
5. Bradford, M.M., 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72: 248-254.
   
6. Bugajska-Schretter, A., A. Pastore, L. Vangelista, H. Rumpold, R. Valenta and S. Spitzauer, 1999. Molecular and immunological characterization of carp parvalbumin, a major fish allergen. Int. Arch. Allergy Immunol., 118: 306-308.
   CrossRef    PMid:10224422    
7. Coffee, C.J. and R.A. Bradshaw, 1973. Carp muscle calcium-binding protein. I. Characterization of the tryptic peptides and the complete amino acid sequence. J. Biol. Chem., 248: 3305-3312.
   PMid:4700462    
8. Elsayed, S., 2002. Fish Allergy and the Codfish Allergen Model on Food Allergy and Intolerance. In: Brostoff, J. and S.J. Challacombe (Eds.), 2nd Edn., Saunders, London, pp: 425-433.
   
9. Elsayed, S. and K. Aas, 1971a. Isolation of purified allergens (cod) by isoelectric focusing. Int. Arch. Allergy Appl. Immunol., 40: 428-438.
   CrossRef    PMid:4993766    
10. Elsayed, S. and K. Aas, 1971b. Characterization of a major allergen (cod): Observation on effect of denaturation on the allergenic activity. J. Allergy, 47: 283-291.
    CrossRef    
11. Elsayed, S. and H. Bennich, 1975. The primary structure of allergen M from cod. Scand. J. Immunol., 4: 203-208.
    CrossRef    PMid:1145128    
12. Elsayed, S. and J. Apold, 1983. Immunochemical analysis of cod fish allergen M: Locations of the immunoglobulin binding sites as demonstrated by the native and synthetic peptides. J. Allergy., 38: 449-459.
    CrossRef    
13. Guo, F., K. Hiroaki and K. Shiomi, 2012. Purification, immunological properties and molecular cloning of two allergenic parvalbumins from the crimson sea bream, Evynnis japonica. Food Chem., 132: 835-840.
    CrossRef    
14. Hamada, Y., H. Tanaka and S. Ishizaki, 2003. Purification, reactivity with IgE and cDNA cloning of parvalbumin as the major allergen of mackerels. Food Chem. Toxicol., 41: 1149-1156.
    CrossRef    
15. Hansen, T.K., C. Bindslev-Jensen, P.S. Skov and L.K. Poulsen, 1997. Codfish allergy in adults: IgE cross-reactivity among fish species. Ann. Allerg. Asthma Im., 78: 187-194.
    CrossRef    
16. Lindstrøm, C.D.V., T. van Dô, I. Hordvik, C. Endresen and S. Elsayed, 1996. Cloning of two distinct cDNAs encoding parvalbumin, the major allergen of Atlantic Salmon (Salmo salar). Scand. J. Immunol., 44: 335-344.
    CrossRef    PMid:8845026    
17. Pascual, C., M.M. Esteban and J.F. Crespo, 1992. Fish allergy: Evaluation of the importance of cross-reactivity. J. Pediatr., 121: 29-34.
    CrossRef    
18. Sampson, H.A., 1999. Food allergy. Part I: Immunopathogenesis and clinical disorders. J. Allergy Clin. Immun., 103: 717-728.
    CrossRef    
19. Sampson, H.A., 2003. Food allergy. J. Allergy Clin. Immun., 111: 540-547.
    CrossRef    
20. Shiomi, K., S. Hayashi, M. Ishikawa, K. Shimakura and Y. Nagashima, 1998. Identification of parvalbumin as an allergen in horse mackerel muscle. Fisheries Sci., 64: 300-304.
    
21. Shiomi, K., Y. Hamada, K. Sekiguchi, K. Shimakura and Y. Nagashima, 1999. Two classes of allergens, parvalbumins and higher molecular weight substances, in Japanese eel and bigeye tuna. Fisheries Sci., 65: 943-948.
    
22. Strehler, E.E., H.M. Eppenberger and C.W. Heizmann, 1977. Isolation and characterization of parvalbumin from chicken leg-muscle. FEBS Lett., 72: 127-133.
    CrossRef    
23. Swoboda, I., A. Bugajska-Schretter, P. Verdino, W. Keller, W.R. Sperr and P. Valent, 2002. Recombinant carp parvalbumin, the major cross-reactive fish allergen: A tool for diagnosis and therapy of fish allergy. J. Immunol., 168: 4576-4584.
    CrossRef    PMid:11971005    
24. Towbin, H., T. Staehelin and J. Gordon, 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. P. Natl. Acad. Sci. USA, 76: 4350-4354.
    CrossRef    PMid:388439 PMCid:PMC411572    
25. Van Do, T., I. Hordvik and C. Endresen, 2005. Characterization of parvalbumin, the major allergen in Alaska Pollack and comparison with codfish Allergen M. Mol. Immunol., 42: 345-353.
    CrossRef    PMid:15589323    
26. Zheng, C., X. Wang, Y. Lu and Y. Liu, 2012. Rapid detection of fish major allergen parvalbumin using super paramagnetic nanoparticle-based lateral flow immunoassay. Food Control, 26: 446-452.
    CrossRef    

Competing interests

The authors have no competing interests.

Open Access Policy

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Copyright

The authors have no competing interests.