Abstract
Chicken serum albumin (CSA) is a hen’s egg yolk allergen causing IgE-mediated allergy. The objective of this study was to produce a recombinant version of CSA and compare its IgE reactivity to natural CSA (nCSA). CSA was cloned and expressed as a soluble fraction in the yeast Kluyveromyces lactis (K. lactis) protein expression system. The gene encoding CSA was amplified with a C-terminal hemagglutinin epitope tag by polymerase chain reaction (PCR) and cloned into the pKLAC2 expression vector prior to transforming into K. lactis. Recombinant CSA (rCSA) was purified by immunoprecipitation. Twenty-one patients allergic to hen’s egg white were examined for sensitisation against nCSA. 38% of patients were found to be sensitised to CSA based on Western immunoassay. Immunoglobulin E (IgE) binding capacity of rCSA and nCSA was analysed by ELISA using sera from patients sensitised to CSA. Levels of IgE-binding were similar for both the recombinant and the natural CSA, indicating the existence of similar epitopes. rCSA produced in this study is a potential candidate to be used in component-resolved diagnosis (CRD) of egg yolk allergy. The usefulness of rCSA in CRD of egg yolk allergy warrants further characterisation using sera from patients with allergy to hen’s egg yolk in future studies.
Highlights
Hen’s (Gallus domesticus) egg is a popular protein source for humans throughout the world.Egg white in relation to food allergy has been well investigated, unlike the egg yolk
The main objective of this study was to produce a recombinant version of Chicken serum albumin (CSA) and characterize its Immunoglobulin E (IgE)-binding capacity in comparison to natural CSA
This study presents the successful production of IgE-reactive Recombinant CSA (rCSA) using K. lactis yeast strain
Summary
Hen’s (Gallus domesticus) egg is a popular protein source for humans throughout the world.Egg white in relation to food allergy has been well investigated, unlike the egg yolk. The study by Okada et al estimated that 9.1% of children with hen’s egg allergy were positive for egg yolk slightly contaminated with egg white, based on oral food challenge (OFC) [2]. In vitro sensitisation to those egg yolk allergens in children diagnosed with egg allergy have been clearly demonstrated in recent studies [5,6]. Understanding of allergy to hen’s egg yolk in the clinical setting is very limited and there is a lack of well-defined reagents for diagnosis of egg yolk allergy. It has been shown that classifying infants with hen’s egg allergy into egg yolk tolerant and egg yolk reactive with slight egg white contamination is useful for predicting the natural course of egg allergy in early childhood [2]. Accurate diagnosis of egg white and egg yolk allergy is crucial in better management of egg allergy and avoiding unnecessary dietary restrictions
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