Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products, National Institutes for Food and Drug Control, No. 31, Huatuo Road, Biomedical Base,Daxing District, Beijing, China.
Luminescence. 2020 Dec;35(8):1408-1415. doi: 10.1002/bio.3905. Epub 2020 Aug 31.
Although enormous success has been achieved with anti-PD-1/PD-L1 and anti-CTLA-4 monoclonal antibodies (mAbs), their unsatisfactory response rate in cancer patients has been driving the research and development of novel immune checkpoint inhibitors (ICIs). Anti-LAG-3 mAbs, as one of the most promising candidates, are now being tested for various human cancers at different stages of clinical trials. Here, we describe the development and validation of a reporter gene assay (RGA) to measure the bioactivity of anti-LAG-3 mAbs. We established the bioassay based on parental Raji cells and a Jurkat cell line stably transfected with human LAG-3 gene and luciferase reporter elements controlled by nuclear factor of activated T cell (NFAT) from the IL-2 promoter. After optimization of key parameters, the established RGA showed excellent precision, specificity, accuracy, and stability. The mechanism of action (MOA) relatedness and the excellent assay performance make the RGA suitable for the characterization, lot release, and stability test of anti-LAG-3 mAbs.
尽管抗 PD-1/PD-L1 和抗 CTLA-4 单克隆抗体（mAbs）取得了巨大成功，但它们在癌症患者中的不尽如人意的反应率促使人们研究和开发新型免疫检查点抑制剂（ICIs）。抗 LAG-3 mAbs 作为最有前途的候选药物之一，目前正在不同临床试验阶段针对各种人类癌症进行测试。在这里，我们描述了一种报告基因检测（RGA）的开发和验证，以测量抗 LAG-3 mAbs 的生物活性。我们基于亲本 Raji 细胞和稳定转染人 LAG-3 基因和受白细胞介素 2（IL-2）启动子核因子活化 T 细胞（NFAT）控制的荧光素酶报告元件的 Jurkat 细胞系建立了生物测定法。在优化了关键参数后，所建立的 RGA 显示出优异的精密度、特异性、准确性和稳定性。作用机制（MOA）相关性和出色的检测性能使 RGA 适用于抗 LAG-3 mAbs 的特征描述、批次放行和稳定性测试。
