Recombinant, Fluorescent, Peptidomimetic Tracer for Immunodetection of Imidaclothiz

文献类型： 外文期刊

第一作者： Ding, Yuan

作者： Ding, Yuan;Hua, Xiude;Du, Mei;Yang, Qian;Hou, Lina;Wang, Limin;Liu, Fengquan;Wang, Minghua;Ding, Yuan;Hua, Xiude;Du, Mei;Yang, Qian;Hou, Lina;Wang, Limin;Liu, Fengquan;Wang, Minghua;Liu, Fengquan;Gonzalez-Sapienza, Gualberto

作者机构：

期刊名称：ANALYTICAL CHEMISTRY （ 影响因子：6.986； 五年影响因子：6.755 ）

ISSN： 0003-2700

年卷期： 2018 年 90 卷 23 期

页码：

收录情况： SCI

摘要： Peptidomimetic and anti-immunocomplex peptides, which can be readily isolated from a phage-display library, have shown great potential for small-molecule immunoassay development because they typically improve the sensitivity and avoid the use of chemical haptens as coatings or tracer antigens. However, phage-borne peptides are unconventional immunoassay reagents, which greatly limits their use in commercial applications, and require secondary reagents for detection. In order to overcome these limitations, we used C2-15, a peptidomimetic of imidaclothiz, as a model peptide fused to emerald-green fluorescent protein (EmGFP) at the N-terminus (C2-15-EmGFP) and C-terminus (EmGFP-C2-15) to generate novel fluorescent-peptide tracers. Both recombinant fluorophores reacted with similar affinity to the anti-imidaclothiz monoclonal antibody 1E(7), but because of its higher expression, C2-15-EmGFP was chosen to develop a competitive Fluorescent peptide tracer magnetic-separation fluorescence immunoassay (MSFIA). After a competitive step with the analyte, the C2-15-EmGFP-antibody complex bound to the magnetic beads was separated with a magnet, and because of the fast dissociation of the peptide-antibody interaction, the fluorescence signal was detected following the spontaneous dissociation of the complex in fresh buffer. The concentration of imidaclothiz causing the 50% inhibitory concentration (IC50) was 11.00 ng mL(-1), and the MSFIA performed with excellent recovery and had a good correlation with high-performance liquid chromatography in different matrices.

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