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

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 developme...

Full description

Saved in:
Bibliographic Details
Published in:Heliyon 2023-06, Vol.9 (6), p.e17401-e17401, Article e17401
Main Authors: Duan, Maoqin, Yu, Chuanfei, Yang, Yalan, Fu, Zhihao, Liu, Chunyu, Du, Jialiang, Li, Meng, Guo, Sha, Yu, XiaoJuan, Xu, Gangling, Mei, Yuting, Wang, Lan
Format: Article
Language:English
Subjects:
Anti-CD25 antibody
Bioassay
IL-2
Reporter gene assay
Citations: Items that this one cites
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:2405-8440
2405-8440
DOI:10.1016/j.heliyon.2023.e17401