Abstract：
In preclinical protein therapeutic development studies, the emergence of anti-drug antibodies (ADA) can potentially impact drug pharmacokinetics and safety. While immunogenicity assessment is not mandatory in preclinical studies, banking samples can be valuable for interpreting unexpected pharmacological responses. Immunoassays that use generic reagents across different drug molecules can simplify ADA assessment and expedite sample evaluations. This work showcases the ability of the Gyrolab automated immunoassay platform to detect and quantify both drug-free and drug-bound (total) ADAs to monoclonal antibody (mAb) therapeutics in cynomolgus monkey preclinical studies. Compared to the previously reported total ADA ELISA, the Gyrolab assay exhibited a wider signal dynamic range and increased drug tolerance. Similar sensitivity, dynamic range and cut point factors were observed for four therapeutic mAbs of different isotypes using the Gyrolab assay. Here we present a comparison of ADA assays using bridging ELISA, total ADA ELISA and total ADA Gyrolab formats in a cynomolgus monkey study where the subjects were treated with a single dose of a mAb therapeutic. We demonstrate that the total ADA assays detected host ADA responses at earlier time points compared to the bridging ELISA. The Gyrolab assay has the best correlation between signal-to-noise (S/N) and titer over a wide ADA concentration range, highlighting the utility of Gyrolab in S/N reporting of ADA response to eliminate the need for secondary titer assays. Collectively, our results demonstrate that the generic ADA Gyrolab assay minimizes the necessity for extensive assay development and optimization for therapeutic mAbs, streamlining preclinical immunogenicity assessment to enable interpretation of pharmacological data.
摘要：
在临床前蛋白质治疗开发研究中，抗药物抗体(ADA)的出现可能影响药物的药代动力学和安全性.虽然免疫原性评估在临床前研究中不是强制性的，银行样本对于解释意外的药理学反应可能是有价值的。使用跨不同药物分子的通用试剂的免疫测定可以简化ADA评估并加快样品评估。这项工作展示了Gyrolab自动免疫测定平台在食蟹猴临床前研究中检测和量化无药和药物结合（总）ADAs对单克隆抗体（mAb）疗法的能力。与以前报道的总ADAELISA相比，Gyrolab分析显示出更宽的信号动态范围和更高的药物耐受性。类似的敏感度，使用Gyrolab测定法观察了四种不同同种型治疗性单克隆抗体的动态范围和截止点因子。在这里，我们提出了使用桥接ELISA的ADA测定的比较，在食蟹猴研究中的总ADAELISA和总ADAGyrolab形式,其中受试者用单剂量的mAb治疗剂治疗。我们证明,与桥接ELISA相比,总ADA测定在较早的时间点检测到宿主ADA应答。Gyrolab测定在很宽的ADA浓度范围内具有信噪比（S/N）和滴度之间的最佳相关性，强调Gyrolab在ADA反应的S/N报告中的实用性，以消除对二次滴度测定的需要。总的来说，我们的结果表明，通用ADAGyrolab测定最大限度地减少了广泛的测定开发和优化治疗性单克隆抗体的必要性，简化临床前免疫原性评估，以实现药理学数据的解释。
