Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Changchun 130062, PR China.
Biosens Bioelectron. 2014 Nov 15;61:241-4. doi: 10.1016/j.bios.2014.05.032. Epub 2014 May 22.
A dual labeled probe was synthesized by coating gold nanoparticles (AuNPs) with anti-κ-CN monoclonal antibody (McAb) and horseradish peroxidase (HRP) enzyme on their surface. The McAb was used as detector and HRP was used as label for signal amplification catalytically oxidize the substrate. AuNPs were used as bridges between the McAb and HRP. Based on the probe, an immunoassay was developed for ultrasensitive detection of κ-CN in bovine milk samples. The assay has a linear response range within 4.2-560 ng mL(-1). The limit of detection (LOD) was 4.2 ng mL(-1) which was 10 times lower than that of traditional McAb-HRP based ELISA. The recoveries of κ-CN from three brand bovine milk samples were from 95.8% to 111.0% that had a good correlation (R(2)=0.998) with those obtained by official standard Kjeldahl method. For higher sensitivity and as simple as the traditional ELISA, the developed immunoassay could provide an alternative approach for ultrasensitive detection of κ-CN in bovine milk sample.
一种双标记探针通过在金纳米粒子（AuNPs）表面涂覆抗κ-CN 单克隆抗体（McAb）和辣根过氧化物酶（HRP）酶来合成。McAb 用作检测器，HRP 用作信号放大的标签，通过催化氧化底物。AuNPs 用作 McAb 和 HRP 之间的桥梁。基于该探针，开发了一种用于检测牛乳中κ-CN 的超灵敏免疫测定法。该测定法在 4.2-560ngmL(-1) 范围内具有线性响应范围。检测限（LOD）为 4.2ngmL(-1)，比传统的 McAb-HRP 基于 ELISA 的检测限低 10 倍。从三个品牌的牛乳样品中回收的 κ-CN 的回收率为 95.8%至 111.0%，与通过官方标准凯氏定氮法获得的回收率具有良好的相关性（R(2)=0.998）。为了提高灵敏度并尽可能简单地模拟传统 ELISA，所开发的免疫测定法可以为牛乳样品中κ-CN 的超灵敏检测提供替代方法。
