J Korean Med Sci.  2024 Feb;39(6):e54. 10.3346/jkms.2024.39.e54.

Variation in the Allergenicity of Scrambled, Boiled, Short-Baked and Long-Baked Egg White Proteins

• Gantulga P
• Lee J
• Jeong K
• Jeon SA
• Lee S

• KMID: 2553305
• DOI: http://doi.org/10.3346/jkms.2024.39.e54

Abstract

Background
Hen’s egg white (HEW) is the most common cause of food allergy in children which induces mild to fatal reactions. The consultation for a proper restriction is important in HEW allergy. We aimed to identify the changes in HEW allergenicity using diverse cooking methods commonly used in Korean dishes.
Methods
Crude extract of raw and 4 types of cooked HEW extracts were produced and used for sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE), enzyme-linked immunosorbent assay (ELISA), and ELISA inhibition assays using 45 serum samples from HEW allergic and tolerant children. Extracts were prepared; scrambled without oil for 20–30 seconds in frying pan without oil, boiled at 100°C for 15 minutes, short-baked at 180°C for 20 minutes, and long-baked at 45°C for 12 hours with a gradual increase in temperature up to 110°C for additional 12 hours, respectively.
Results
In SDS-PAGE, the intensity of bands of 50–54 kDa decreased by boiling and baking. All bands almost disappeared in long-baked eggs. The intensity of the ovalbumin (OVA) immunoglobulin E (IgE) bands did not change after scrambling; however, an evident decrease was observed in boiled egg white (EW). In contrast, ovomucoid (OM) IgE bands were darker and wider after scrambling and boiling. The IgE binding reactivity to all EW allergens were weakened in short-baked EW and considerably diminished in long-baked EW. In individual ELISA analysis using OVA⁺OM⁺ serum samples, the median of specific IgE optical density values was 0.435 in raw EW, 0.476 in scrambled EW, and 0.487 in boiled EW. Conversely, it was significantly decreased in short-baked (0.406) and long-baked EW (0.012). Significant inhibition was observed by four inhibitors such as raw, scrambled, boiled and short-baked HEW, but there was no significant inhibition by long-baked HEW (IC₅₀ > 100 mg/mL).
Conclusion
We identified minimally reduced allergenicity in scrambled EW and extensively decreased allergenicity in long-baked EW comparing to boiled and short-baked EW as well as raw EW. By applying the results of this study, we would be able to provide safer dietary guidence with higher quality to egg allergic children.

Keyword

Egg White Proteins; Food Allergy; Cooking; Allergenicity

Figure

• 

  Fig. 1 SDS-PAGE profiles of raw and 4 different cooked hen’s egg white proteins. M-marker: 1, Raw EW; 2, Scrambled EW; 3, Boiled EW; 4, Short-baked EW; and 5, Long-baked EW.SDS-PAGE = sodium dodecyl-sulfate polyacrylamide gel electrophoresis, EW = egg white.

• 

  Fig. 2 Immunoblot analysis of raw and 4 different cooked hen’s egg white proteins with pooled serum samples of patients. (A) OVA+OM+-pooled serum samples. (B) OVA+OM−-pooled serum samples. (C) OVA−OM−-pooled serum samples. M-marker: 1, Raw EW; 2, Scrambled EW; 3, Boiled EW; 4, Short-baked EW; 5, Long-baked EW.OVA = ovalbumin, OM = ovomucoid, EW = egg white.

• 

  Fig. 3 Individual ELISA of raw and 4 different cooked hen’s egg white crude extracts with individual serum samples of patients. (A) OVA+OM+ serum samples of 15 patients. (B) OVA+OM− serum samples of 15 patients. (C) OVA−OM− serum samples of 15 patients. Results are expressed by median and range of OD 450 value.ELISA = enzyme-linked immunosorbent assay, OVA = ovalbumin, OM = ovomucoid, OD = optical density.*P < 0.05, **P < 0.01, ***P < 0.001.

• 

  Fig. 4 Egg white inhibition ELISA of raw and 4 different cooked hen’s egg white crude extract with OVA+OM+-pooled serum samples of patients. Inhibitors: Raw EW, Scrambled EW, Boiled EW, Short-Baked EW, Long-baked EW and FBS.ELISA = enzyme-linked immunosorbent assay, OVA = ovalbumin, OM = ovomucoid, EW = egg white, FBS = fetal bovine serum, IC50 = half-maximal inhibitory concentration.

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