Purification of Plasmid DNA by Multimodal Chromatography

A Rita Silva-Santos; Sara Sousa Rosa; Duarte Miguel F Prazeres; Ana M Azevedo

doi:10.1007/978-1-0716-0872-2_10

Purification of Plasmid DNA by Multimodal Chromatography

Methods Mol Biol. 2021:2197:193-205. doi: 10.1007/978-1-0716-0872-2_10.

Authors

A Rita Silva-Santos¹, Sara Sousa Rosa¹, Duarte Miguel F Prazeres¹, Ana M Azevedo²

Affiliations

¹ Department of Bioengineering, Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.
² Department of Bioengineering, Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal. a.azevedo@tecnico.ulisboa.pt.

PMID: 32827138
DOI: 10.1007/978-1-0716-0872-2_10

Abstract

Multimodal (MM) chromatography can be described as a chromatographic method that uses more than one mode of interaction between the target molecule and the ligand to achieve a particular separation. Owing to its advantages over traditional chromatography, such as higher selectivity and capacity, its application for the purification of biomolecules with therapeutic interest has been widely studied. The potential of MM chromatography for the purification of plasmid DNA has been demonstrated. In this chapter, a downstream process for the purification of supercoiled plasmid DNA using MM chromatography with two different ligands-Capto™ adhere and PPA HyperCell™-is described. In both the cases, the purification process yields a high purity and highly homogeneous sc plasmid product.

Keywords: Downstream processing; Multimodal chromatography; Plasmid DNA; Purification; Supercoiling.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Chromatography / methods*
DNA, Circular / isolation & purification*
DNA, Superhelical / isolation & purification
Dialysis
Escherichia coli / genetics
Plasmids / isolation & purification*

Substances

DNA, Circular
DNA, Superhelical