计量学报2025，Vol.46Issue(5)：754-761,8.DOI:10.3969/j.issn.1000-1158.2025.05.19
中国仓鼠卵巢细胞宿主DNA残留检测标准物质
Chinese Hamster Ovary Cell Genome DNA Residue Reference Material for Testing
摘要
Abstract
To solve the traceability problem of the exogenous DNA residue determination method(qPCR method)in Chinese Pharmacopoeia(2020 edition),a large-scale cultivation of Chinese hamster ovary(CHO)cells was conducted.Various techniques such as column chromatography,ethanol precipitation,and ultrafiltration were employed to extract high-purity CHO genomic DNA.Digital PCR,which has been extensively studied and optimized,was chosen as the quantitative method for genomic DNA.The results showed that the digital PCR-based method effectively overcame matrix interference issues and demonstrated better consistency than traditional UV detection.The uniformity and stability of the standard substance were verified,and the copy number concentration was determined using 5 different digital PCR platforms from 10 units,resulting in a final value of(1.06±0.12)×104 copies/μL.This standard substance can facilitate the development of DNA residual testing kits and promote mutual recognition and standardization of DNA residual testing results among biopharmaceutical companies,ensuring traceability of the testing results.关键词
生物计量学/中国仓鼠卵巢细胞/宿主DNA/残留检测/数字PCR/标准物质Key words
biometrology/CHO cell/genome DNA/residual detection/digital PCR/reference materials分类
通用工业技术引用本文复制引用
梁文,曹梅霞,刘刚..中国仓鼠卵巢细胞宿主DNA残留检测标准物质[J].计量学报,2025,46(5):754-761,8.基金项目
国家市场监督管理总局科技计划(2022MK024) （2022MK024）
上海市科技创新行动(23002400200) （23002400200）
国家质量基础设施体系专项(2021YFF0600705) （2021YFF0600705）
