Development and Evaluation of a Reporter Gene Assay for DKK-1 Antibody Drugs Based on the Wnt/β-Catenin Pathway
ZHAO Xiang
HOU Sheng
XU Jin
ZHANG Dapeng
FU Rongrong
XU Mengjiao
WANG Fugui
QIAN Weizhu
LI Jun
Abstract：The determination of bioactivity is crucial for ensuring the safety and efficacy of therapeutic anti-bodies,necessitating an accurate reflection of the drug's mechanism of action(MOA).DKK-1 antibodies,as novel targeted therapies,have shown significant potential in the treatment of osteoporosis and cancer.However,the current bioactivity evaluation for DKK-1 antibodies primarily relies on the detection of alka-line phosphatase activity.This process is complex,time-consuming,and highly variable,which limits the development and quality control of DKK-1 antibodies.To establish a high-performance bioactivity assay for DKK-1 antibodies,this study introduced a reporter gene assay(RGA).By transfecting HEK 293 cells with a reporter gene plasmid stably expressing luciferase reporter under the control of TCF/LEF response elements,we successfully established a reporter gene assay based on the DKK-1 antibody MOA—the Wnt/β-catenin pathway.Through optimization and simulation experiment evaluation,the method has shown promising potential and reliability in practical application.The development of this innovative ap-proach is expected to enhance the screening,assessment,and quality control of DKK-1 antibody drugs in future development.
Keywords：bioactivityantibody drugsDKK-1reporter gene assay
Publication Date：2024-10-28
Online Publishing Date：2026-08-28(First online date of this platform, not the publication date of the document)
Pages：8( 183-190 )