Poor patient adherence to antiretroviral medication represents a major obstacle for managing disease and reducing rates of new HIV infections. The measurement of patient drug levels is the most objective method of determining adherence. Tenofovir and tenofovir diphosphate are metabolites of some of the most common HIV medications for treatment and prevention and can be quantified by mass spectrometry. Here, we report the development of a competitive enzyme linked immunoassay as a simplified approach for detecting tenofovir and tenofovir diphosphate. Monoclonal antibodies were produced by two tenofovir-hapten conjugates and screened for binding to immobilized tenofovir, and then for competition by tenofovir and tenofovir diphosphate. Antibody specificity was evaluated against adenosine phosphates, which are close structural analogs. We performed numerical simulations of reaction equilibrium to guide assay optimization. When used to evaluate spiked tenofovir in plasma and spiked tenofovir diphosphate in red blood cell lysate, the optimized assay had high sensitivity and specificity.

中文翻译：

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HIV药物代谢物Tenofovir和Tenofovir Diphosphate的免疫分析。

患者对抗逆转录病毒药物的依从性差是控制疾病和降低新的HIV感染率的主要障碍。测量患者药物水平是确定依从性的最客观方法。替诺福韦和替诺福韦二磷酸酯是一些用于治疗和预防的最常见的HIV药物的代谢产物，可以通过质谱定量。在这里，我们报告竞争性酶联免疫测定法的发展，作为检测替诺福韦和替诺福韦二磷酸的简化方法。由两种替诺福韦-半抗原缀合物产生单克隆抗体，并筛选与固定化替诺福韦的结合，然后筛选与替诺福韦和替诺福韦二磷酸的竞争。评估了针对磷酸腺苷的抗体特异性，磷酸腺苷是紧密的结构类似物。我们进行了反应平衡的数值模拟，以指导测定的优化。当用于评估血浆中加标的替诺福韦和红细胞裂解液中加标的替诺福韦二磷酸时，优化后的测定具有很高的灵敏度和特异性。