Abstract：
E6011, a humanized anti-fractalkine monoclonal antibody, is under development for the treatment of various inflammatory diseases, such as rheumatoid arthritis. Therapeutic antibodies may induce production of anti-drug antibodies (ADA) that may deteriorate efficacy and/or enhance immunogenic reaction. It is important to have an ADA assay to understand the characteristics of biotherapeutics under development. A simple and reproducible assay has thus been developed for the determination of ADA against E6011 in monkey and human serum by electrochemiluminescence (ECL) detection. An immune-complex of biotinylated E6011, ADA, and ruthenium-labeled E6011 was attached to avidin-coated wells for ECL signal detection. Screening and confirmatory cutpoints were determined to judge negative or positive ADA. Sensitivity of ADA was 1.61 and 1.34 ng/mL in monkey and human serum, respectively. Accuracy and precision of the assay were within ±20% and 20%, respectively. Drug tolerance of the assay in monkey and human sera was ensured up to 100 and 1000 μg/mL E6011 at the surrogate ADA levels of 1 and 4 μg/mL, respectively. The developed assay was successfully applied to ADA quantification in monkeys and humans in support of immunogenicity assessments.
摘要：
E6011，人源化抗Fractalkine单克隆抗体，正在开发用于治疗各种炎症性疾病，如类风湿性关节炎。治疗性抗体可诱导抗药物抗体(ADA)的产生,其可降低功效和/或增强免疫原性反应。重要的是要进行ADA测定以了解正在开发的生物治疗剂的特征。因此，已开发出一种简单且可重复的测定法，用于通过电化学发光（ECL）检测来测定猴子和人血清中针对E6011的ADA。生物素化E6011，ADA的免疫复合物，将钌标记的E6011连接到抗生物素蛋白包被的孔上用于ECL信号检测。确定筛选和确认切点以判断阴性或阳性ADA。ADA在猴和人血清中的敏感性为1.61和1.34ng/mL，分别。测定的准确度和精密度在±20%和20%以内，分别。在1和4μg/mL的替代ADA水平下，确保了猴子和人血清中测定的药物耐受性高达100和1000μg/mLE6011，分别。所开发的测定成功地应用于猴和人中的ADA定量以支持免疫原性评估。
