Darwish Ibrahim A, Ali Mohammad A H, Alsalhi Mohammed S, Zhang Daohong
Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University P.O. Box 2457 Riyadh 11451 Saudi Arabia
College of Food Science and Engineering, Northwest A&F University Yangling 712100 Shaanxi China.
RSC Adv. 2024 Mar 11;14(12):8167-8177. doi: 10.1039/d4ra00202d. eCollection 2024 Mar 6.
This study describes, for the first time, the development and validation of a novel ultrasensitive chemiluminescence enzyme immunoassay (CLEIA) for the quantification of atezolizumab (ATZ), a monoclonal antibody approved by the FDA for treatment of different types of cancer. The assay involved the non-competitive binding of ATZ to its specific antigen (PD-L1 protein). The immune complex of PD-L1/ATZ formed on the internal surface of the plate wells was quantified by a novel chemiluminescence (CL)-producing horseradish peroxidase (HRP) reaction. The reaction employed a highly efficient CL enhancer for the HRP-luminol-hydrogen peroxide reaction which was 4-(imidazol-1-yl)phenol. The conditions of the CLEIA and its detection system were refined, and the optimum procedures were established. The CLEIA was validated in accordance with the guidelines of immunoassay validation for bioanalysis, and all the validation criteria were acceptable. The assay's limit of detection and limit of quantitation were 12.5 and 37.5 pg mL, respectively, with a working dynamic range of 25-800 pg mL. The assay enables the accurate and precise quantitation of ATZ in human plasma samples without any interferences from endogenous substances and/or the plasma matrix. The results of the proposed CLEIA were favourably comparable with those of a pre-validated enzyme-linked immunosorbent assay using a colorimetric detection system. The CLEIA is characterized by simple and high throughput features. The CLEIA is superior to the existing analytical methodologies for ATZ. The proposed CLEIA has a great value in the quantitation of ATZ in clinical settings for assessment of its pharmacokinetics, therapeutic drug monitoring, and refining the safety profile.
本研究首次描述了一种新型超灵敏化学发光酶免疫分析法（CLEIA）的开发与验证，该方法用于定量测定阿特珠单抗（ATZ），这是一种经美国食品药品监督管理局（FDA）批准用于治疗不同类型癌症的单克隆抗体。该分析方法涉及ATZ与其特异性抗原（程序性死亡配体1蛋白，PD-L1）的非竞争性结合。通过一种新型的产生化学发光（CL）的辣根过氧化物酶（HRP）反应，对在板孔内表面形成的PD-L1/ATZ免疫复合物进行定量。该反应采用了一种高效的CL增强剂用于HRP-鲁米诺-过氧化氢反应，即4-（咪唑-1-基）苯酚。对CLEIA及其检测系统的条件进行了优化，并确定了最佳程序。CLEIA按照生物分析免疫分析验证指南进行了验证，所有验证标准均符合要求。该分析方法的检测限和定量限分别为12.5和37.5 pg/mL，工作动态范围为25 - 800 pg/mL。该分析方法能够准确、精确地定量测定人血浆样本中的ATZ，不受内源性物质和/或血浆基质的干扰。所提出的CLEIA的结果与使用比色检测系统的预先验证的酶联免疫吸附测定法的结果具有良好的可比性。CLEIA具有操作简单和高通量的特点。CLEIA优于现有的ATZ分析方法。所提出的CLEIA在临床环境中对ATZ进行定量以评估其药代动力学、治疗药物监测和完善安全性方面具有重要价值。
