Abstract：
Monitoring anti-drug antibodies (ADAs) to infliximab and adalimumab is critical to treatment management in various autoimmune disorders. The growing need for proactive therapeutic monitoring further requires the detection of ADAs in the presence of measurable concentrations of infliximab or adalimumab. To provide robust analytical assays for clinical application, we evaluated two automated immunoassays developed using ImmunoCAP™ technology and based on the bridging format to measure serum ADAs to infliximab and adalimumab respectively. Without an acid-dissociation step, these research prototype assays can detect a positive control monoclonal ADA towards infliximab and adalimumab, ranging from < 25 ng/ml to 10,000 ng/mL. Both assays exhibit imprecision less than 20% at different ADA titer levels and can distinguish ADAs towards different drug targets. In method comparison using authentic patient samples, the quantitative results of the ADA assays are not directly comparable to two existing clinical immunoassays for ADAs (correlation coefficient rs = 0.673 for infliximab ADAs; rs = 0.510 for adalimumab ADAs), presumably due to the lack of commutable ADA standards and the polyclonal nature of ADAs. Nevertheless, there is qualitative agreement between the methods when evaluating putative positive and negative patient samples (overall agreement 0.83 for infliximab ADAs; 0.76 for adalimumab ADAs). Biotin and high levels of rheumatoid factors may interfere with the performance of the automated assays due to competitive binding with the biotinylated drug and non-specific formation of bridging complexes. The two ImmunoCAP assays can provide new analytical methods for proactive therapeutic monitoring of adalimumab and infliximab.
摘要：
监测英夫利昔单抗和阿达木单抗的抗药物抗体（ADAs）对于各种自身免疫性疾病的治疗管理至关重要。对主动治疗监测的日益增长的需求进一步需要在可测量浓度的英夫利昔单抗或阿达木单抗存在下检测ADMA。为临床应用提供强大的分析检测，我们评估了使用ImmunoCAP™技术和基于桥接格式分别测量英夫利昔单抗和阿达木单抗血清ADAs的两种自动化免疫测定法.没有酸解离步骤，这些研究原型试验可以检测英夫利昔单抗和阿达木单抗的阳性对照单克隆ADA，范围从<25ng/ml到10,000ng/mL。两种测定在不同的ADA滴度水平下表现出小于20%的不精确性,并且可以区分针对不同药物靶标的ADA。在使用真实患者样本的方法比较中，ADA测定的定量结果与两种现有的ADAs临床免疫测定不能直接比较（英夫利昔单抗ADAs的相关系数rs=0.673；阿达木单抗ADAs的相关系数rs=0.510），可能是由于缺乏可交换的ADA标准和ADA的多克隆性质。然而，在评估推定的阳性和阴性患者样本时,方法之间存在定性一致性(英夫利昔单抗ADAs的总体一致性为0.83;阿达木单抗ADAs的总体一致性为0.76).由于与生物素化药物的竞争性结合和桥接复合物的非特异性形成,生物素和高水平的类风湿因子可能干扰自动化测定的性能。两种ImmunoCAP测定法可以为阿达木单抗和英夫利昔单抗的主动治疗监测提供新的分析方法。
