Advancing recombinant protein production in CHO cells through metabolic engineering.

Chinese hamster ovary (CHO) cells serve as the predominant platform for producing recombinant therapeutic proteins in biopharmaceutical manufacturing, the production capacity of which relies heavily on efficient protein synthesis, folding, and secretion pathways. However, during high-density and prolonged cultivation, these cells frequently encounter bottlenecks-including excessive lactate and ammonia accumulation, redox imbalance, and endoplasmic reticulum (ER) stress-which ultimately constrain both the yield and quality of target protein. To overcome these limitations, metabolic engineering has emerged as a key strategy; through systematic modification of the CHO cellular metabolic network, it enhances recombinant protein yield, optimizes critical product qualities such as glycosylation, and improves overall process robustness. This review summarizes recent advances in CHO cell metabolic engineering, encompassing the regulation of central metabolic pathways, glycosylation engineering, cell cycle and metabolic reprogramming, culture condition optimization, byproduct accumulation control, and the application of systems biology and artificial intelligence technologies, including genome-scale metabolic modeling, machine learning-guided target prediction, and dynamic process control. These advances have significantly reduced biopharmaceutical production costs, improved scalability, and shortened time-to-market for monoclonal antibodies and other complex biologics. As the field transitions from single-gene manipulation toward multi-target, dynamic, and system-level rational design, metabolic engineering is advancing CHO cells into more efficient and intelligent "cell factories", thereby providing sustained momentum for the industrial production of biologics.