Journal article
Hypoallergenic variant of the major egg white allergen gal d 1 produced by disruption of cysteine bridges
P Dhanapala, D Withanage-Dona, MLK Tang, T Doran, C Suphioglu
Nutrients | MDPI | Published : 2017
DOI: 10.3390/nu9020171
Abstract
Background: Gal d 1 (ovomucoid) is the dominant allergen in the chicken egg white. Hypoallergenic variants of this allergen can be used in immunotherapy as an egg allergy treatment approach. We hypothesised that disruption of two of the nine cysteine-cysteine bridges by site-directed mutagenesis will allow the production of a hypoallergenic variant of the protein; Methods: Two cysteine residues at C192 and C210 in domain III of the protein were mutated to alanine using site-directed mutagenesis, to disrupt two separate cysteine-cysteine bridges. The mutated and non-mutated proteins were expressed in Escherichia coli (E. coli) by induction with isopropyl β-D-1-thiogalactopyranoside (IPTG). Th..
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Funding Acknowledgements
We would like to thank the Poultry Cooperative Research Centre (CRC) (established and supported under the Australian Government's Cooperative Research Centres Program) and Deakin University's Molecular and Medical Research (MMR) Strategic Research Centre (SRC) for providing this study with the required research funding, the Australian Animal Health Laboratory (AAHL) of Commonwealth Scientific and Industrial Research Organisation (CSIRO) for supplying animal tissues required for the study, and the Murdoch Childrens Research Institute (MCRI) at the Royal Children's Hospital Melbourne for supplying egg-allergic patients' sera which was crucial for the immunological analysis. The Murdoch Childrens Research Institute is supported by the Victorian Government's Operational Infrastructure Support Program. Author P.D. was supported by a Deakin University Post Graduate Research Scholarship and a Poultry CRC Top-Up PhD Scholarship.