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

doi:10.1016/j.heliyon.2023.e17401

Establishing a novel and sensitive assay for bioactivity determination of anti-CD25 antibodies

Heliyon. 2023 Jun 24;9(6):e17401. doi: 10.1016/j.heliyon.2023.e17401. eCollection 2023 Jun.

Authors

Maoqin Duan¹, Chuanfei Yu¹, Yalan Yang¹, Zhihao Fu¹, Chunyu Liu¹, Jialiang Du¹, Meng Li¹, Sha Guo¹, XiaoJuan Yu¹, Gangling Xu¹, Yuting Mei¹, Lan Wang¹

Affiliation

¹ 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.

Abstract

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.

Keywords: Anti-CD25 antibody; Bioassay; IL-2; Reporter gene assay.