Establishment and validation of a reporter gene assay to determine the bioactivity of CpG QCX1 adjuvant in vitro
GE Jun
HE Peng
JIANG Chuan-wen
HUANG Hong-ying
ZHANG Shuang
LU Jin-biao
LI Jian-qiang
HU Zhong-yu
Abstract：Objective:To establish and validate a reporter gene assay for in vitro determining the bioactivity of CpG QCX1 adjuvant.Methods:Following the guidelines of ICH Q2(R2)and the General Chapter 9401 of the Chinese Pharmacopoeia 2020 edition,volume Ⅲ,combined with design of experiment(DOE),the analytical method was optimized and its specificity,relative accuracy,intermediate precision,linearity,and range were validated.Results:The concentration range of the test sample was determined to be 0.195～50 μg·mL-1,dilution factor of 1∶2,cell concentration of 4.5×105 cells·mL-1,and incubation time of 16 hours.The method could specifically detect CpG QCX1,with a linear range of 50%～185%,relative bias of-2.7%to-1.0%for five activity levels,geometric coefficient of variation less than 10%,and a linear regression correlation coefficient(r)of 0.996,all of which met the requirements.Conclusion:The method established in this study has good specificity,accuracy,robustness,and durability,and can be used for the quality control of related CpG products.
Keywords：CpG QCX1in vitro biological activitymethod establishmentmethod validationquality control
Publication Date：2025-08-03
Online Publishing Date：2026-08-14(First online date of this platform, not the publication date of the document)
Pages：7( 1748-1754 )