Large Scale Printing of Robust HPLC Medium via Layer-by-Layer Stereolithography
Hanrong Wen, H. Lu, Zhuoheng Zhou, Kaiyue Sun, Yinjia Huang, Juxing Zeng, Yuchen Wang, Lianzhong Luo, Xu Chen, Jianzhong Xu, Xin Zhang, Xiaofei Wang, Sebastiaan Eeltink, Bo Zhang
Abstract
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.