通过使用荧光酶标仪和KinExA 3200免疫传感器开发两种不同形式的阿法替尼生物分析的异源荧光免疫测定。,Scientific Reports

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通过使用荧光酶标仪和KinExA 3200免疫传感器开发两种不同形式的阿法替尼生物分析的异源荧光免疫测定。
Scientific Reports ( IF 3.8 ) Pub Date : 2019-10-14 , DOI: 10.1038/s41598-019-51288-5
Ibrahim A Darwish ₁ , Haitham AlRabiah ₁ , Mohammed A Hamidaddin _{1,

2}

Affiliation

阿法替尼（AFT）是一种有效且高度选择性的药物，用于治疗各种实体瘤，包括非小细胞肺癌（NSCLC）。为了确保安全有效地治疗AFT癌症患者，应监测其血浆浓度。因此，需要灵敏的免疫测定来测量血浆样品中的AFT浓度。在这项研究中，开发了两种不同形式的异源荧光免疫测定法，并通过了AFT生物分析验证。这些测定是使用荧光板读数器的基于微孔的荧光免疫测定（FIA）和使用KinExA 3200免疫传感器的动力学排斥测定（KinExA）。FIA和KinExA都是使用相同的试剂开发的：小鼠抗AFT抗体，与牛血清白蛋白（AFT-BSA）偶联的固相固定化AFT，以及用异硫氰酸荧光素（FITC-IgG）标记的山羊抗小鼠IgG来产生信号。对这两种测定的分析性能进行了比较评估，结果表明，尽管两种测定具有可比的准确性，但KinExA在灵敏度和精密度方面均优于FIA。此外，FIA和KinExA都是用于AFT生物分析的现有色谱方法的更好替代品。预计拟议的FIA和KinExA将在确保AFT在临床环境中安全有效的治疗方面做出有效贡献。对于现有的AFT生物分析色谱方法，FIA和KinExA都是更好的选择。预计拟议的FIA和KinExA将在确保AFT在临床环境中安全有效的治疗方面做出有效贡献。对于现有的AFT生物分析色谱方法，FIA和KinExA都是更好的选择。预计拟议的FIA和KinExA将在确保AFT在临床环境中安全有效的治疗方面做出有效贡献。

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Development of two different formats of heterogeneous fluorescence immunoassay for bioanalysis of afatinib by employing fluorescence plate reader and KinExA 3200 immunosensor.

Afatinib (AFT) is a potent and highly selective drug used to treat various solid tumors including non-small cell lung cancer (NSCLC). To ensure safe and effective treatment of cancer patients with AFT, its plasma concentrations should be monitored. Thus, sensitive immunoassays are required for measuring AFT concentrations in plasma samples. In this study, two different formats of heterogeneous fluorescent immunoassays were developed and validated for AFT bioanalysis. These assays were microwell-based fluorescence immunoassay (FIA) using fluorescence plate reader and kinetic exclusion assay (KinExA) using KinExA 3200 immunosensor. Both FIA and KinExA were developed using the same reagents: mouse anti-AFT antibody, solid-phase immobilized AFT conjugated with bovine serum albumin protein (AFT-BSA), and goat anti-mouse IgG labelled with fluorescein isothiocyanate (FITC-IgG) for signal generation. The analytical performances of both assays were comparatively evaluated, and the results revealed that although both assays had comparable accuracies, KinExA was superior to FIA in terms of sensitivity and precisions. Moreover, both FIA and KinExA were better alternatives to the existing chromatographic methods for bioanalysis of AFT. The proposed FIA and KinExA are anticipated to effectively contribute in ensuring safe and effective treatment with AFT in clinical settings.

更新日期：2019-10-14

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