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

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.