Bottom-up Precision Engineering of Porous Medium for High-Performance Chromatography Based on Stereolithography 3D Printing

Engineering of high-performance chromatography medium, has long been esteemed 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 bottom-up engineering strategy for chromatography media utilizing 3D-printing technology. Combining micrometer stereolithography and nanometer pore chemistry, the strategy enables precision engineering of high-performance separation media in diverse scales, shapes and chemistry. Between 1,000 devices, exceptional performance consistency was demonstrated by superb column-to-column and batch-to-batch reproducibility (retention time CV 2.04%). Ultra-fast separations of intact proteins and antibodies were realized in reversed-phase and hydrophobic interaction chromatography: within 1 minute, resolution > 1.5 was achieved. Purification of native proteins was directly amplified three orders of magnitude: 12 mg hemeproteins were isolated in 8 min at negligible scaling-up cost, supporting liter-scale processing within 7 h on one 20 mm i.d. column. With unprecedented advantages in fully automatic and parallel production capacity, high-fidelity microstructure and chemistry across dimensions, and highly efficient method transfer and scaling-up, the stereolithography engineered chromatography media may open a green and sustainable path to speeding up separation and purification processes from primary analysis to mass-production of biomolecular entities, as demanded in biosynthesis and pharmaceutical industries.