化学
NFAT公司
单克隆抗体
生物制药
报告基因
抗体
细胞培养
分子生物学
荧光素酶
Jurkat细胞
生物测定
计算生物学
基因
免疫系统
生物化学
T细胞
基因表达
生物
免疫学
转染
转录因子
遗传学
作者
Lan Wang,Chuanfei Yu,Yalan Yang,Kai Gao,Junzhi Wang
标识
DOI:10.1016/j.jpba.2017.05.011
摘要
Being regarded as the ‘cancer panacea’, the anti-PD-1/anti-PD-L1 monoclonal antibodies (mAbs) have become the R&D focus of biopharmaceutical industries. Several marketed such mAbs have been proved particularly effective in treating various cancers. However, the cell-based bioassay to measure the biological activities of the anti-PD-1/anti-PD-L1 mAbs as the lot release or stability test has been a great challenge to quality control laboratories due to the immunomodulating nature of the mAbs. Here, we describe the development and validation of a reporter gene assay consisting of two-cell systems to measure the bioactivity of the anti-PD-1/anti-PD-L1 mAbs. We have generated two cell lines, the CHO-PD-L1-CD3L cell line that stably expresses PD-L1 and the membrane-anchored anti-CD3 single chain antibody fragment (scFv) named CD3L and the Jurkat-PD-1-NFAT cell line that stably expresses PD-1 and the luciferase gene under the control of the NFAT response elements from the IL-2 promoter. The results show good dose-dependent responsiveness to the mAbs and excellent performance characteristics including specificity, accuracy and precision. The biological relevance of the assay, the passage stability of the two cell lines, and the capability of measuring various anti-PD-1/anti-PD-L1 mAbs render this assay applicable not only in lot release and stability test but also in characterization and development of new anti-PD1/anti-PD-L1 mAbs.
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