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Recent advancements in site\u2010specific integration\u2010based (SSI) platforms in Chinese hamster ovary (CHO) cells can mitigate the high transcriptional variation observed with random integration and the low industrial relevance of transient expression by providing a flexible platform for mAb expression from a consistent clonal background. This work applies a novel SSI\u2010based expression system capable of generating isogenic cell pools in less than 1 month to systematically compare the expression of ten sequence variants of two therapeutically relevant mAbs from two genomic loci under industrially relevant culture conditions. Eight single amino acid mutations in trastuzumab resulted in reduced productivity compared to the wild\u2010type mAb in batch cultures, and three mutations maintained a low\u2010expressing phenotype in fed\u2010batch cultures. The mutations resulted in variant\u2010specific patterns of decreased domain stability and increased ER stress. The application of industrially relevant SSI systems in developability workflows could strengthen the understanding of the sequence determinants of mAb expression to improve mAb design, candidate selection, and process development decisions.<\/jats:p>","DOI":"10.1002\/biot.70102","type":"journal-article","created":{"date-parts":[[2025,8,18]],"date-time":"2025-08-18T04:47:03Z","timestamp":1755492423000},"source":"Crossref","is-referenced-by-count":1,"title":["Evaluation of \u201cDifficult\u2010to\u2010Express\u201d Monoclonal Antibodies in a CHO\u2010Based Hybrid Site\u2010Specific Integration System Under Industrially Relevant Conditions"],"prefix":"10.1002","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9071-9363","authenticated-orcid":false,"given":"Alana C.","family":"Szkodny","sequence":"first","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering University of Delaware Newark Delaware 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