Tirzepatide, a dual GIP and GLP-1 receptor agonist, has emerged as one of the most successful therapeutic peptides for the treatment of type 2 diabetes and obesity. The global demand has expanded to the hundreds of kilograms and is projected to exceed one ton annually. However, the large-scale manufacturing remains challenging due to the structural complexity and the presence of closely related impurities arising from peptide synthesis. This is particularly highlighted by the limitations of conventional preparative chromatography, where yield and productivity are compromised by the intrinsic purity–yield trade-off. Therefore, efficient downstream processing ensuring scalability and process economy are essential for sustainable supply of therapeutic peptides.
In this study, we developed a highly efficient two-step approach for tirzepatide purification utilizing YMC-Triart Prep stationary phases. YMC-Triart Prep stationary phases featuring excellent mechanical and chemical durability offer various functionalities (C18, C8, etc.) leading to diverse separation characteristics. Our systematic method screening by YMC-Triart Prep stationary phases, pH values, and organic solvents contributed to the development of the optimal combinations. For example, in the first step, Triart Prep Phenyl-S under neutral conditions (mobile phase: 20 mM ammonium acetate aqueous solution with acetonitrile) effectively removed major impurities while maintaining a yield close to 98%. In the second step, Triart Prep C4-S under basic conditions (mobile phase: 50 mM ammonium bicarbonate aqueous solution with acetonitrile) further increased purity to >99%, demonstrating the advantage of YMC-Triart Prep stationary phases with high durability even under basic conditions.
To overcome the purity–yield trade-off in the second step, twin-column continuous chromatography (MCSGP-AutoPeak) was employed. MCSGP-AutoPeak ensures reproducibility between preparative fractions by automatically recycling low-purity side fractions containing the target compound and compensating for retention time variations based on chromatograms. Compared with conventional purification, which provided approximately 70% recovery in this study, MCSGP-AutoPeak achieved 95% recovery even at fivefold higher loading, leading to doubled productivity while maintaining >99% purity. This result indicates that the large-scale purification with MCSGP-AutoPeak using two 800 mmI.D. columns projected that annual tirzepatide production exceeding one ton is feasible. Notably, our downstream processing as shown above achieves high yield (recovery), significantly reducing the required amount of crude peptide and further emphasizing its economic advantage.
In conclusion, utilizing YMC-Triart Prep stationary phases and twin-column continuous chromatography (MCSGP-AutoPeak) provides a powerful, scalable, and cost-efficient downstream processing for not only tirzepatide but also the other peptides. Our approach in this study should enable meeting industrial-scale requirements while ensuring regulatory-compliant purity and yield.