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A key element of a robust TI CLD system is the construction of a high\u2010performance TI host cell line, which significantly influences the productivity, product quality, and cell line stability of expressed biologics. In this study, we implemented a low\u2010copy transfection strategy by optimizing transfection dosages to efficiently achieve over 30% single\u2010copy integrants in a single transfection. This approach dramatically reduces TI host cell screening efforts, enabling rapid isolation of functional TI host cells. In addition, using mRNA transfection of Cre recombinase enabled faster pool recovery and eliminated concerns related to residual Cre coding sequence integration. Under non\u2010optimized fed\u2010batch conditions, this resulting TI system supported monoclonal antibody (mAb) titers of up to 8.03\u00a0g\/L in 15\u2010L bioreactors.<\/jats:p>","DOI":"10.1002\/biot.70165","type":"journal-article","created":{"date-parts":[[2025,12,11]],"date-time":"2025-12-11T15:10:53Z","timestamp":1765465853000},"source":"Crossref","is-referenced-by-count":2,"title":["Leveraging Transfection Dose Streamlines the Establishment of a Chinese Hamster Ovary Cell Targeted Integration System"],"prefix":"10.1002","volume":"20","author":[{"given":"Tao","family":"Sun","sequence":"first","affiliation":[{"name":"Cell Line Development WuXi Biologics  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Zhou","sequence":"additional","affiliation":[{"name":"Cell Line Development WuXi Biologics  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Zhang","sequence":"additional","affiliation":[{"name":"Cell Line Development WuXi Biologics  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nan","family":"Yan","sequence":"additional","affiliation":[{"name":"Cell Line Development WuXi Biologics  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ding","family":"Huang","sequence":"additional","affiliation":[{"name":"Cell Culture Process Development WuXi Biologics  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kee Wee","family":"Tan","sequence":"additional","affiliation":[{"name":"Cell Line Development WuXi Biologics  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sam","family":"Zhang","sequence":"additional","affiliation":[{"name":"Microbial and Viral Platform WuXi Biologics  Hangzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hang","family":"Zhou","sequence":"additional","affiliation":[{"name":"Bioprocess Research &amp; 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