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- Title
Immunological Comparison of Native and Recombinant Hen’s Egg Yolk Allergen, Chicken Serum Albumin (Gal d 5), Produced in Kluveromyces lactis.
- Authors
King, Samuel; Suphioglu, Cenk; De Silva, Chamika; Dhanapala, Pathum; Doran, Timothy; Tang, Mimi
- 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 <italic>Kluyveromyces lactis</italic> (<italic>K. lactis</italic>) protein expression system. The gene encoding CSA was amplified with a <italic>C</italic>-terminal hemagglutinin epitope tag by polymerase chain reaction (PCR) and cloned into the pKLAC2 expression vector prior to transforming into <italic>K. lactis</italic>. 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.
- Subjects
DIAGNOSIS of food allergies; ALLERGIES; EGGS; ALLERGENS; ENZYME-linked immunosorbent assay; GENETIC techniques; IMMUNOLOGICAL adjuvants; POLYMERASE chain reaction; RECOMBINANT proteins; SERUM albumin; WESTERN immunoblotting; YEAST; PRECIPITIN tests; DIAGNOSIS
- Publication
Nutrients, 2018, Vol 10, Issue 6, p757
- ISSN
2072-6643
- Publication type
Article
- DOI
10.3390/nu10060757