Establishing a novel and sensitive assay for bioactivity determination of anti-CD25 antibodies
Maoqin Duan,
Chuanfei Yu,
Yalan Yang,
Zhihao Fu,
Chunyu Liu,
Jialiang Du,
Meng Li,
Sha Guo,
XiaoJuan Yu,
Gangling Xu,
Yuting Mei,
Lan Wang
Affiliations
Maoqin Duan
Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech, Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, Beijing 102629, China
Chuanfei Yu
Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech, Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, Beijing 102629, China
Yalan Yang
Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech, Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, Beijing 102629, China
Zhihao Fu
Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech, Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, Beijing 102629, China
Chunyu Liu
Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech, Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, Beijing 102629, China
Jialiang Du
Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech, Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, Beijing 102629, China
Meng Li
Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech, Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, Beijing 102629, China
Sha Guo
Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech, Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, Beijing 102629, China
XiaoJuan Yu
Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech, Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, Beijing 102629, China
Gangling Xu
Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech, Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, Beijing 102629, China
Yuting Mei
Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech, Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, Beijing 102629, China
Lan Wang
Corresponding author. Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, 31# HUATUO Road, Beijing 102629, China.; Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech, Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, Beijing 102629, China
Anti-CD25 antibodies have been approved for renal transplantation and has been used prior to and during transplantation by the Food and Drug Administration (FDA). However, no reported bioassays have been reflected the mechanism of action (MOA) of anti-CD25 antibodies. Here, we describe the development and validation of a reporter gene assay (RGA) based on the engineered C8166-STAT5RE-Luc cells expressing endogenous IL-2 receptors and a STAT5-inducible element-driven firefly luciferase in C8166 cell lines. The RGA was fully validated according to the International Conference on the Harmonization of Technical Requirements for the Registration of Pharmaceuticals for the Human Use-Q2 (ICH-Q2). After optimization, the assay showed excellent specificity, linearity, accuracy, precision, and robustness. Due to the MOA relatedness and the excellent assay performance, the RGA is suitable for exploring the critical quality attributes (CQAs), release inspection, comparability and stability of anti-CD25 mAbs.
