Liu Xiao-Zhi, Zhang Bin, Zhao Wei, Li Geng, Zhou Mei-Ling, Wei Jing-Shuang, Zhou Jing-Hua, Gao Jian, Wang Zhi-Ming
State Key Laboratory of Antibody Research, Development, New Drug Research and Development Company Ltd., North China PharmaceuticalCorporation, Shijiazhuang, 050015, China.
Key Laboratory for Family Planning and Healthy Birth, National Health and Family Planning Commision, Hebei Research Institute for Family Planning, Shijiazhuang, 050071, China.
Biologicals. 2019 Nov;62:65-71. doi: 10.1016/j.biologicals.2019.09.004. Epub 2019 Sep 18.
The residual DNA derived from host cells in antibody drugs have potential safety risks. In this paper, the antibody in the test sample was removed by magnetic bead separation method, and the residual DNA were quantitatively determined by Q-PCR method. The residual DNA in the sample was analyzed according to the standard curve. We validated the species specificity, accuracy, precision, quantitative restrictions, reproducibility of this method. The results showed the linearrange was of 1 × 10～1 × 10 pg/μL and the curve linear was good, this method can specifically detect CHO cell DNA. Compared with the method of extracting residual DNA by magnetic beads, the method has the advantages of simplicity, rapidity and low cost, and can be used for quantitative determination of the residual host cell DNA in antibody drugs producted by CHO cells.
抗体药物中源自宿主细胞的残留DNA存在潜在安全风险。本文采用磁珠分离法去除测试样品中的抗体，并用Q-PCR法对残留DNA进行定量测定。根据标准曲线对样品中的残留DNA进行分析。我们验证了该方法的物种特异性、准确性、精密度、定量限、重现性。结果表明，线性范围为1×10～1×10 pg/μL，曲线线性良好，该方法能特异性检测CHO细胞DNA。与磁珠提取残留DNA的方法相比，该方法具有简便、快速、成本低的优点，可用于CHO细胞生产的抗体药物中残留宿主细胞DNA的定量测定。
