Biocatalysis & Biotransformation Conference

Presentation abstract:

Immobilized enzyme reactors (IMERs) offer a powerful strategy to simplify and accelerate nucleic acid processing by enabling enzymatic reactions directly on chromatographic supports. In this study, we explore monolith-based IMERs for catalytic transformations of nucleic acids in bioprocessing. Alongside restriction enzymes for plasmid linearization, a range of nucleic acid – degrading enzymes – including DNases and RNases – were examined in both analytical and preparative workflows to demonstrate the platform’s versatility.
Chromatographic and electrophoretic analyses confirmed that enzyme immobilization preserved catalytic performance and reaction specificity. Restriction enzyme IMERs achieved complete plasmid linearization suitable for in vitro transcription (IVT) without the need for heat inactivation or enzyme removal. RNase IMERs enabled selective RNA degradation for quantification of residual DNA in IVT mixtures, while specialized RNase IMERs facilitated targeted RNA digestion, supporting applications in RNA integrity and stability analysis. Conversely, DNase IMERs provided controlled DNA digestion after IVT and in HEK293 cell lysates, demonstrating their utility in both analytical and preparative processes.
Altogether, these results underscore the versatility of monolith-based IMERs as robust, reusable, and application-specific tools for nucleic acid processing and analysis. Their operational simplicity, flow-through design, and compatibility with standard chromatography systems position them as valuable components of integrated, automated bioprocessing and analytical platforms for RNA, DNA, and gene therapy manufacturing.