Structure-guided sodium butyrate analogs enhance antibody productivity in CHO cells via metabolic and cell-cycle...

Chinese hamster ovary (CHO) cells are the primary platform for therapeutic antibody production. Although histone deacetylase inhibitors such as sodium butyrate (NaBu) can enhance recombinant expression, their growth-inhibitory and cytotoxic effects often limit volumetric productivity. Here, a structure-guided docking strategy was applied to prioritize NaBu-derived small-molecule additives (SMAs) for experimental screening in CHO antibody-producing cell lines. A lead combination (D1 +D4) increased volumetric antibody titers by approximately 2-3 fold while maintaining high cell viability (>95%) under the tested conditions. Cell-based analyses indicated reduced apoptotic markers and altered cell-cycle distributions in D1 +D4-treated cultures relative to NaBu. Metabolite profiling further revealed reduced by-product accumulation, including lactate and ammonia, together with a distinct intracellular energy and redox state compared with control cultures. Importantly, N-glycan profiles and charge variants of the produced antibodies remained comparable between control and D1 +D4-treated cultures. The productivity benefit of D1 +D4 was further maintained in bench-scale stirred-tank and wave bioreactor systems, supporting process relevance. Together, these results demonstrate that structure-guided NaBu analog screening can identify practical additives that enhance CHO antibody productivity with minimal impact on product quality.