利用鲨鱼源单域抗体荧光免疫层析法快速检测鱼糜中鸭蛋卵白蛋白（D-OVA）

IF 9.8 1区农林科学 Q1 CHEMISTRY, APPLIED

Food Chemistry Pub Date : 2025-06-10 DOI:10.1016/j.foodchem.2025.145117

Zhong-Lin Li , Ling-Jing Zhang , Kai-Yuan Cao , Jian-Lian Huang , Chang-Gong Zhang , Le-Chang Sun , Yu-Lei Chen , Min-Jie Cao

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引用次数: 0

摘要

建立了一种基于鲨鱼源单域抗体的荧光免疫层析试纸条（FITS），用于鱼糜制品中主要食物过敏原鸭蛋清蛋白（D-OVA）的现场检测。D-OVA采用离子交换色谱法纯化，质谱法鉴定。特异性可变新抗原受体（VNAR）是一种从白斑黄萎病病菌噬菌体展示VNAR合成抗体文库中筛选出的新抗原受体。单域抗体在HEK293F细胞中表达并纯化。Western blot和生物层干涉（BLI）分析表明，单结构域抗体14A2和14A8对D-OVA具有特异性和亲和力。基于竞争荧光免疫层析分析，将量子点（QDs）与抗体14A2偶联，形成D-OVA-FITS。开发的D-OVA-FITS的检测限（LOD）为0.04 μg/mL， IC₅₀为0.3 μg/mL，检测时间为15 min。总体而言，D-OVA- fits具有准确性和特异性，适用于鱼糜产品中D-OVA的快速定量检测。

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Rapid detection of duck egg ovalbumin (D-OVA) in surimi via fluorescent immunochromatographic assay utilizing shark-derived single-domain antibody

A fluorescent immunochromatographic test strip (FITS) based on shark-derived single-domain antibodies for on-site detection the major food allergen of duck egg ovalbumin (D-OVA) in surimi products was developed. D-OVA was purified using ion-exchange chromatography and confirmed by mass spectrometry. Specific variable new antigen receptor (VNAR) was selected from a synthetic antibody library derived from phage-displayed VNARs of Chiloscyllium plagiosum. Single-domain antibodies were expressed in HEK293F cells and purified. Western blot and Bio-Layer Interferometry (BLI) analyses demonstrated that single-domain antibodies 14A2 and 14A8 exhibited both specificity and affinity for D-OVA. Quantum dots (QDs) were conjugated to antibody 14A2 to develop D-OVA-FITS, based on a competitive fluorescent immunochromatographic assay. The developed D-OVA-FITS revealed a limit of detection (LOD) of 0.04 μg/mL, an IC₅₀ of 0.3 μg/mL, with detection time of 15 min. Overall, the D-OVA-FITS exhibited accuracy, and specificity and it is applicable to detect D-OVA in surimi products rapidly and quantitatively.

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来源期刊

Food Chemistry 工程技术-食品科技

CiteScore

16.30

自引率

10.20%

发文量

3130

审稿时长

122 days

期刊介绍： Food Chemistry publishes original research papers dealing with the advancement of the chemistry and biochemistry of foods or the analytical methods/ approach used. All papers should focus on the novelty of the research carried out.