Enhancing TGF-β1 production in CHO cells via MS-275 treatment, Bax/Bak knockout, and PACEsol overexpression

Abstract Efficient production of recombinant human transforming growth factor-β1 (rhTGF-β1) in Chinese hamster ovary (CHO) cells is limited by low yields and inefficient precursor processing. Here, we developed an integrated strategy combining chemical and genetic engineering. Screening of additives revealed that the histone deacetylase inhibitor MS-275 increased specific productivity ( q hTGF−β1 ) but inhibited growth, without affecting precursor processing. To counteract this, Bax and Bak double knockout cells were generated, conferring apoptosis resistance and enabling higher rhTGF-β1 yields under MS-275 treatment. Further enhancement was achieved by overexpressing soluble furin (PACEsol), which improved precursor maturation. Together, these interventions synergistically increased mature rhTGF-β1 production up to 8.8 µg/mL, representing a ~ 2.5-fold increase compared with the parental line. These findings highlight that simultaneously targeting apoptosis resistance and precursor processing can effectively overcome key bottlenecks in CHO cell culture and substantially improve rhTGF-β1 production.