Abstract
Minicircles are non-viral delivery vectors with promising features for biopharmaceutical applications. These vectors are plasmid-derived circular DNA molecules that are obtained in vivo in Escherichia coli by the intramolecular recombination of a parental plasmid, which generates a minicircle containing the eukaryotic therapeutic cassette of interest and a miniplasmid containing the prokaryotic backbone. The production process results thus in a complex mixture, which hinders the isolation of minicircle molecules from other DNA molecules. Several strategies have been proposed over the years to meet the challenge of purifying and obtaining high quality minicircles in compliance with the regulatory guidelines for therapeutic use. In minicircle purification, the characteristics of the strain and parental plasmid used have a high impact and strongly affect the purification strategy that can be applied. This review summarizes the different methods developed so far, focusing not only on the purification method itself but also on its dependence on the upstream production strategy used.
Highlights
Biopharmaceuticals are classically defined as a pharmaceutical drug with active agents of biological origin that are manufactured by biotechnological processes
The goal of this review is to address the several methodologies used for minicircle purification and their dependence on the selected upstream production strategy
This review provides an overview of processes used to purify minicircle vectors, with a special focus on the strategies used to separate minicircles from miniplasmids and parental plasmids
Summary
Biopharmaceuticals are classically defined as a pharmaceutical drug with active agents of biological origin that are manufactured by biotechnological processes. One of the main advantages of biopharmaceutical products is their higher specificity and activity in comparison to conventional drugs. The field has been marked by a continuous increase in the number and types of biopharmaceuticals developed and marketed to treat diseases such as cancer, inflammation-related conditions, hemophilia and diabetes. Biopharmaceuticals encompass a wide range of products such as antibodies (which currently hold the highest number of product approvals), recombinant enzymes, cell-based therapies and nucleic-acid based products, among others. In the area of nucleic acid based products, several types are reported, namely antisense oligonucleotides, aptamers, modified RNA molecules, small interfering RNAs, gene therapy vectors and DNA or RNA vaccines (Walsh 2018). Broad and comprehensive analysis of the progress and developments on the nucleic acid biopharmaceutical field can be found in recent reviews (e.g., Prazeres and Monteiro 2014; Uludag et al, 2019)
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