Regeneron Pharmaceuticals, Bioanalytical Sciences, 777 Old Saw Mill River Rd, Tarrytown, New York, 10591, USA.
Pfizer, 401 N Middletown Rd, Pearl River, New York, 10965, USA.
AAPS J. 2021 Oct 4;23(6):109. doi: 10.1208/s12248-021-00643-4.
Monoclonal antibodies (mAbs) are a leading class of biotherapeutics. In oncology, patients often fail on early lines of biologic therapy to a specific target. Some patients may then enroll in a new clinical trial with a mAb specific for the same target. Therefore, immunoassays designed to quantify the current mAb therapy or assess immunogenicity to the drug may be susceptible to cross-reactivity or interference with residual prior biologics. The impact of two approved anti-PD-1 mAbs, pembrolizumab and nivolumab, was tested in several immunoassays for cemiplimab, another approved anti-PD-1 mAb. The methods included a target-capture drug concentration assay, a bridging anti-drug antibody (ADA) assay and a competitive ligand-binding neutralizing antibody (NAb) assay. We also tested bioanalytical strategies to mitigate cross-reactivity or interference in these assays from other anti-PD-1 biologics. Both pembrolizumab and nivolumab cross-reacted in the cemiplimab drug concentration assay. This was mitigated by addition of antibodies specific to pembrolizumab or nivolumab. ADA specific for pembrolizumab and nivolumab did not interfere in the cemiplimab ADA assay. However, pembrolizumab and nivolumab generated a false-positive response in a target-capture NAb assay. Our results demonstrate that similar exogenous pre-existing anti-PD-1 mAbs (biotherapeutics) such as pembrolizumab and nivolumab are detected and accurately quantified in the cemiplimab drug concentration assay. However, once steady state is achieved for the new therapy, prior biologics would likely not be detected. Cross-reactivity and interference in immunoassays from previous treatment with class-specific biotherapeutic(s) pose significant bioanalytical challenges, especially in immuno-oncology.
单克隆抗体（mAbs）是一类主要的生物治疗药物。在肿瘤学中，患者经常在针对特定靶点的早期生物治疗中失败。一些患者可能会参加针对同一靶点的新临床试验，使用针对特定靶点的 mAb。因此，用于定量当前 mAb 治疗或评估对药物免疫原性的免疫测定法可能容易受到交叉反应或与残留先前生物制剂的干扰。两种已批准的抗 PD-1 mAb（pembrolizumab 和 nivolumab）在几种用于另一种已批准的抗 PD-1 mAb cemiplimab 的免疫测定法中进行了测试。这些方法包括靶向捕获药物浓度测定法、桥接抗药物抗体（ADA）测定法和竞争配体结合中和抗体（NAb）测定法。我们还测试了生物分析策略，以减轻这些测定法中来自其他抗 PD-1 生物制剂的交叉反应或干扰。pembrolizumab 和 nivolumab 在 cemiplimab 药物浓度测定法中均发生交叉反应。通过添加针对 pembrolizumab 或 nivolumab 的抗体可以减轻这种交叉反应。针对 pembrolizumab 和 nivolumab 的 ADA 不会干扰 cemiplimab ADA 测定法。然而，pembrolizumab 和 nivolumab 在靶向捕获 NAb 测定法中产生了假阳性反应。我们的结果表明，类似的外源性预先存在的抗 PD-1 mAb（生物制剂），如 pembrolizumab 和 nivolumab，在 cemiplimab 药物浓度测定法中被检测到并准确定量。然而，一旦新疗法达到稳态，先前的生物制剂可能不会被检测到。来自先前使用特定类别生物制剂治疗的免疫测定法中的交叉反应和干扰带来了重大的生物分析挑战，尤其是在免疫肿瘤学中。
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