NeuroAllergy Research Laboratory (NARL), School of Life and Environmental Sciences, Faculty of Science, Engineering and Built Environment, Deakin University, 75 Pigdons Road, Geelong, Victoria 3216, Australia.
Poultry Cooperative Research Centre, P.O. Box U242, University of New England, Armidale, NSW 2351, Australia.
Nutrients. 2018 Jun 12;10(6):757. doi: 10.3390/nu10060757.
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 () protein expression system. The gene encoding CSA was amplified with a -terminal hemagglutinin epitope tag by polymerase chain reaction (PCR) and cloned into the pKLAC2 expression vector prior to transforming into . 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.
鸡血清白蛋白（CSA）是一种引起 IgE 介导过敏的母鸡卵黄过敏原。本研究的目的是生产 CSA 的重组版本，并比较其与天然 CSA（nCSA）的 IgE 反应性。CSA 通过聚合酶链反应（PCR）进行克隆和表达为酵母（）蛋白表达系统中的可溶性部分。通过 PCR 扩增 CSA 基因，在其 N 端添加一个血凝素表位标签，并克隆到 pKLAC2 表达载体中，然后转化到酵母中。通过免疫沉淀法纯化重组 CSA（rCSA）。对 21 名对蛋清过敏的患者进行了 nCSA 致敏检测。根据 Western 免疫印迹法，38%的患者被发现对 CSA 过敏。使用对 CSA 敏感的患者的血清通过 ELISA 分析 rCSA 和 nCSA 的 IgE 结合能力。重组和天然 CSA 的 IgE 结合水平相似，表明存在相似的表位。本研究中生产的 rCSA 是卵黄过敏的成分分辨诊断（CRD）的潜在候选物。rCSA 在卵黄过敏的 CRD 中的用途需要进一步通过未来对鸡蛋黄过敏患者的血清进行特征描述来确定。
