Fully automated minicolumn chromatography

Abstract

Miniaturized chromatography columns (minicolumns) operated by automated liquid handlers are an integral part of bioprocess purification development. However, these systems can be limited in both their efficiency and accessibility. Because the minicolumn chromatography operation itself is higher throughput, the lower throughput pre- and post-operation activities become the bottleneck of the workflow. Additionally, method writing and operation of the systems while varying multiple parameters, using a design of experiments approach for example, can be error-prone and resource intensive. Here, we have developed a fully automated minicolumn chromatography system to both address these bottlenecks and improve the accessibility of these systems by allowing users to enter chromatography-relevant information through a simplified user interface. Methods have been developed to automate buffer preparation and protein solution titration leveraging modeling and integrated pH probes with feedback control. Chromatogram generation and fraction pooling has additionally been automated to improve the efficiency of post-chromatography operations. We have also demonstrated the flexibility of the system through an example run where both bind-and-elute chromatography and flowthrough chromatography experiments were performed in parallel. Additionally, all methodology and parameters to operate the system have been shared. We hope this will help interested parties improve the efficiency and accessibility of their minicolumn chromatography systems.