A Facile and Reproducible Method for the Purification of Peptide- and Protein-Functionalized DNA Nanostructures.

Abstract

DNA nanotechnology has emerged as a promising field for biomedical applications, in both the therapeutic and diagnostic domains. The ability of DNA nanostructures to carry cargos in precise numbers and orientations makes them competitive candidates for drug delivery, biosensors, or imaging agents. Two of the main challenges for translating DNA nanostructures from the laboratory to the clinic are achieving cost-effective large-scale production and establishing comprehensive safety profiles. Having the ability to reliably and efficiently purify functionalized DNA nanostructures is key to both challenges and an open question in the field of DNA nanotechnology. Here we present a scalable method for the fast and efficient purification of a high concentration of peptide- or protein-functionalized DNA nanostructures. We use a gravity-driven size exclusion chromatography approach that has the potential to purify DNA nanostructures within 10 min in yields of up to 93% with purities of over 99.9% and is appropriate for both protein and peptide conjugates.