单克隆抗体
中国仓鼠卵巢细胞
基因
细胞生物学
细胞培养
效价
直线(几何图形)
细胞生长
计算机科学
计算生物学
化学
单克隆
生物
细胞
分子生物学
转录组
基因传递
一致性(知识库)
基因表达
转染
生物信息学
遗传增强
作者
Wanjun Lan,Xiaoli Yang,Juanjuan Yao,Lung-JR Lin,Wenzheng Li,Weihai Chen,J Li,Qi Shen,Kang Li,Huiling Li,Yujia Chen,Liang Chen,Kingsley Leung
标识
DOI:10.1186/s13036-025-00610-z
摘要
Chinese hamster ovary (CHO) cell is a widely used cell line for the production of therapeutic proteins. Customarily, CHO host cell line is established through random integration, which requires multiple rounds of screening to identify the optimal producer. In contrast, site-specific integration (SSI) technology can boost cell line development efficiency by directing gene of interest (GOI) to certain genomic loci that supports sustained and stable expression. In this study, by utilizing AI algorithms and robotic systems, we have identified several CHO host cell lines carrying marker gene by Bxb1-mediated SSI technology. After the substitution of marker gene with different types of GOI at specific sites, some of cases delivered a titer exceeding 15 g/L. Thanks to the uniformity of SSI cell lines, protein titer and quality of monoclonal cells can be predicted based on the performance of corresponding minipools. Different monoclonal cells originated from the same minipool also showed consistent protein quality. Furthermore, it was observed that the monoclonal cells obtained by SSI demonstrated consistency in protein titer, quality and genetic stability after 26 consecutive passages (approximately 90 days). In summary, we have successfully developed an effective platform for the construction of SSI cell lines, which allows the rapid and efficient expression of various proteins while maintaining consistency in stability and product quality.
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