Allergy Asthma Immunol Res.  2013 Jan;5(1):42-47. 10.4168/aair.2013.5.1.42.

The Influence of the Presence of Wheat Flour on the Antigenic Activities of Egg White Proteins

• Shin M
• Lee J
• Ahn K
• Lee SI
• Han Y

• KMID: 2167005
• DOI: http://doi.org/10.4168/aair.2013.5.1.42

Abstract

PURPOSE
It is known that ovomucoid, an egg allergen, is heat resistant and remains soluble after heating. However, a recent study showed that the antigenic activity of ovomucoid could be reduced by heating when egg white (EW) was mixed with wheat flour. This study was performed to determine the influence of wheat flour on the antigenic activities of EW proteins when EW is heated, and the influence of the duration of heat treatment.
METHODS
A mixture of EW and wheat flour was kneaded for 10 minutes and then baked at 180degrees C for 10 minutes and 30 minutes. The EW without wheat flour was also heated at 180degrees C for 10 minutes and 30 minutes. The proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and IgE immunoblotting was performed with the pooled sera of 5 egg-allergic patients. The antigenic activities of ovomucoid in different EW samples were measured by inhibition enzyme-linked immunosorbent assay (ELISA).
RESULTS
1) SDS-PAGE: the intensity of the 37-50 kD bands (overlapped bands of ovomucoid and ovalbumin) decreased significantly in the mixture of EW and wheat flour baked for 30 minutes, compared with the mixture baked for 10 minutes, heated EW and raw EW. 2) IgE immunoblot: in the mixture of EW and wheat, a remarkable decrease of IgE reactivity to 37-50 kD was observed when baked for 30 minutes. 3) Inhibition ELISA: the antigenic activity of ovomucoid decreased significantly in the mixture of EW and wheat baked for 30 minutes, but not in the heated pure EW.
CONCLUSIONS
This study showed that the antigenic activity of ovomucoid can be reduced by baking EW with wheat flour. The decrease in ovomucoid antigenicity in the baked mixture of EW and wheat flour was dependent on the time of heat treatment, indicating that heating should be prolonged to achieve a reduction in ovomucoid antigenic activity.

Keyword

Egg white; wheat flour; hot temperature; ovomucoid

MeSH Terms

Acrylic Resins
Egg Proteins
Egg White
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Flour
Heating
Hot Temperature
Humans
Immunoblotting
Immunoglobulin E
Ovomucin
Ovum
Proteins
Sodium Dodecyl Sulfate
Triticum
Acrylic Resins
Egg Proteins
Immunoglobulin E
Ovomucin
Proteins
Sodium Dodecyl Sulfate

Figure

• 

  Fig. 1 SDS-PAGE analysis of heat treated egg white (EW) proteins and EW mixed with wheat flour proteins. Lane 1; Protein marker Lane 2; raw EW, Lane 3; baked EW for 10 min at 180℃, Lane 4; baked EW for 30 min at 180℃, Lane 5; baked EW mixed with wheat flour for 10 min at 180℃, Lane 6; baked EW mixed with wheat flour for 30 min at 180℃.

• 

  Fig. 2 IgE Immunoblotting analysis with pooled sera of egg-allergic patients. (A) shows an immunoblot result using native serum. (B) is a immunoblot result using the pooled sera of egg-allergic patients. Lane 1: Protein marker; Lane 2: raw egg white (EW); Lane 3: baked EW for 10 min at 180℃; Lane 4: baked EW for 30 min at 180℃; Lane 5: baked EW mixed with wheat flour for 10 min at 180℃ and Lane 6: baked EW mixed with wheat flour for 30 min at 180℃.

• 

  Fig. 3 Inhibition enzyme-linked immunosorbent assay analysis for heat treated egg white (EW) and EW mixed with wheat flour proteins for egg-allergic patients' sera.

Reference

1. Bock SA, Atkins FM. Patterns of food hypersensitivity during sixteen years of double-blind, placebo-controlled food challenges. J Pediatr. 1990. 117:561–567.

2. Boyano-Martínez T, García-Ara C, Díaz-Pena JM, Martín-Esteban M. Prediction of tolerance on the basis of quantification of egg white-specific IgE antibodies in children with egg allergy. J Allergy Clin Immunol. 2002. 110:304–309.

3. Eggesbø M, Botten G, Halvorsen R, Magnus P. The prevalence of allergy to egg: a population-based study in young children. Allergy. 2001. 56:403–411.

4. Sicherer SH, Sampson HA. 9. Food allergy. J Allergy Clin Immunol. 2006. 117:S470–S475.

5. Niggemann B, Sielaff B, Beyer K, Binder C, Wahn U. Outcome of double-blind, placebo-controlled food challenge tests in 107 children with atopic dermatitis. Clin Exp Allergy. 1999. 29:91–96.

6. Sampson HA, McCaskill CC. Food hypersensitivity and atopic dermatitis: evaluation of 113 patients. J Pediatr. 1985. 107:669–675.

7. Jung JA, Lee JS, Lee SI. Serum specific IgE to egg White, cow's milk, soybean in the children with atopic dermatitis. Pediatr Allergy Respir Dis. 2003. 13:255–262.

8. Wood RA. The natural history of food allergy. Pediatrics. 2003. 111:1631–1637.

9. Savage JH, Matsui EC, Skripak JM, Wood RA. The natural history of egg allergy. J Allergy Clin Immunol. 2007. 120:1413–1417.

10. Coombs RR, McLaughlan P. Allergenicity of food proteins and its possible modification. Ann Allergy. 1984. 53:592–596.

11. Davis PJ, Williams SC. Protein modification by thermal processing. Allergy. 1998. 53:102–105.

12. Mine Y, Zhang JW. Comparative studies on antigenicity and allergenicity of native and denatured egg white proteins. J Agric Food Chem. 2002. 50:2679–2683.

13. Peng HJ, Chang ZN, Tsai LC, Su SN, Shen HD, Chang CH. Heat denaturation of egg-white proteins abrogates the induction of oral tolerance of specific Th2 immune responses in mice. Scand J Immunol. 1998. 48:491–496.

14. Takagi K, Teshima R, Okunuki H, Sawada J. Comparative study of in vitro digestibility of food proteins and effect of preheating on the digestion. Biol Pharm Bull. 2003. 26:969–973.

15. Bernhisel-Broadbent J, Dintzis HM, Dintzis RZ, Sampson HA. Allergenicity and antigenicity of chicken egg ovomucoid (Gal d III) compared with ovalbumin (Gal d I) in children with egg allergy and in mice. J Allergy Clin Immunol. 1994. 93:1047–1059.

16. Urisu A, Ando H, Morita Y, Wada E, Yasaki T, Yamada K, Komada K, Torii S, Goto M, Wakamatsu T. Allergenic activity of heated and ovomucoid-depleted egg white. J Allergy Clin Immunol. 1997. 100:171–176.

17. Cooke SK, Sampson HA. Allergenic properties of ovomucoid in man. J Immunol. 1997. 159:2026–2032.

18. Kato Y, Watanabe H, Matsuda T. Ovomucoid rendered insoluble by heating with wheat gluten but not with milk casein. Biosci Biotechnol Biochem. 2000. 64:198–201.

19. Shek LP, Soderstrom L, Ahlstedt S, Beyer K, Sampson HA. Determination of food specific IgE levels over time can predict the development of tolerance in cow's milk and hen's egg allergy. J Allergy Clin Immunol. 2004. 114:387–391.

20. Takagi K, Teshima R, Okunuki H, Itoh S, Kawasaki N, Kawanishi T, Hayakawa T, Kohno Y, Urisu A, Sawada J. Kinetic analysis of pepsin digestion of chicken egg white ovomucoid and allergenic potential of pepsin fragments. Int Arch Allergy Immunol. 2005. 136:23–32.

21. Thomas K, Herouet-Guicheney C, Ladics G, Bannon G, Cockburn A, Crevel R, Fitzpatrick J, Mills C, Privalle L, Vieths S. Evaluating the effect of food processing on the potential human allergenicity of novel proteins: international workshop report. Food Chem Toxicol. 2007. 45:1116–1122.

22. Nowak-Wegrzyn A, Fiocchi A. Rare, medium, or well done? The effect of heating and food matrix on food protein allergenicity. Curr Opin Allergy Clin Immunol. 2009. 9:234–237.

23. Kato Y, Oozawa E, Matsuda T. Decrease in antigenic and allergenic potentials of ovomucoid by heating in the presence of wheat flour: dependence on wheat variety and intermolecular disulfide bridges. J Agric Food Chem. 2001. 49:3661–3665.

24. Lemon-Mulé H, Sampson HA, Sicherer SH, Shreffler WG, Noone S, Nowak-Wegrzyn A. Immunologic changes in children with egg allergy ingesting extensively heated egg. J Allergy Clin Immunol. 2008. 122:977–983.