The manufacture of high-performance liquid chromatography (HPLC) medium has long been viewed as an art rather than science
this raised a great challenge in securing separation consistency, method transferability, and scaling-up in purification of biomolecules. Herein, we report a large scale layer-by-layer manufacturing strategy for a high performance chromatography medium utilizing 3D-printing technology. Combining stereolithography 3D printing and porogenic chemistry, the strategy enables parallel production of high-performance separation medium in diverse scales, shapes, and throughput. Between 1,000 printed devices, high performance consistency was demonstrated by column-to-column and batch-to-batch reproducibility (coefficient of variation of retention time, 2.04%). Fast separations of intact proteins were realized in reversed-phase chromatography: within 1 min, resolution >
1.5 was achieved, and nondenatured antibody separation was realized in hydrophobic interaction chromatography. Purification of native proteins was directly amplified by 3 orders of magnitude: 12 mg of hemeproteins was isolated in 8 min at negligible scaling-up cost, supporting liter-scale processing of fermentation within 7 h on one 20 mm i.d. printed column. With advantages in automatic and parallel production capacity, high-fidelity microstructure across dimensions, and highly efficient method transfer and scaling-up, the stereolithographically printed high performance chromatography medium may open a new path to speeding up separation and purification processes from primary analysis to mass-purification of biomolecular entities, as demanded in the biosynthesis and pharmaceutical industries.