Biomedical application of plasmid DNA in gene therapy: A new challenge for chromatography

Abstract

Gene therapy and DNA vaccination are clinical fields gradually emerging in the last few decades, in particular after the discovery of some gene-related diseases. The increased relevance of biomedical applications of plasmid DNA (pDNA) to induce therapeutic effects has had a great impact on biopharmaceutical research and industry. Although there are several steps involved in the pDNA manufacturing process, the several unit operations must be designed and integrated into a global process. After the plasmid has been designed according to the requirements for clinical administeration to humans, it is biosynthesised mainly by an E. coli host. The overriding priority of the production process is to improve plasmid quantity -the production conditions need to be optimised to guarantee pDNA stability and biological activity.The complexity and diversity of biomolecules present on the pDNA-containing extracts represent the main concern and limitation to achieve pure and biologically active pDNA. There has been a recent intenstification of the improvement of existing purification procedures or the establishment of novel schemes for plasmid purification.

Polyphenol‐Containing Nanoparticles: Synthesis, Properties, and Therapeutic Delivery

“…Plasmid DNA (pDNA) is circular DNA which can be genetically engineered to deliver therapeutic genes to target sites. [189,190] Cationic macromolecules are widely used materials to condense pDNA into polyelectrolyte nanocomplexes, which can be easily internalized by cells. The positively charged polymer can form a positive complex with the negatively charged gene and the complex can be adsorbed onto the cell surface by electrostatic attraction.…”

Segregated growth kinetics of Escherichia coli DH5α‐NH36 in exponential‐fed perfusion culture for pDNA vaccine production

“…The clinical application of plasmid DNA (pDNA) is progressing, and commercial availability for vaccination and gene therapy purposes is approaching toward reality [1,2]. DNA vaccines allow foreign genes to be transiently expressed in transfected cells, mimicking intracellular pathogenic infection and triggering both humoral and cellular immune responses [3].…”

A new microfluidic platform for the highly reproducible preparation of non-viral gene delivery complexes

“…11 Considering these drawbacks and because of the clinical quest for an ever-larger quantity of nanocomplexes, the development of tools for scaling up and standardizing the production of gene delivery particles cannot be delayed any longer. 1,[12][13][14][15][16] Microfluidic systems are notable for predictable fluid behavior and are ideal tools for controlling the physical interaction of reagents at the microscale. 17 As microfluidics allows the most accurate manipulation of small amounts of fluids by simply tuning the experimental parameters such as species concentrations and flow rates, this technology has thus been extensively used to produce optimally tailored nanoparticles.…”

“…11 Considering these drawbacks and because of the clinical quest for an ever-larger quantity of nanocomplexes, the development of tools for scaling up and standardizing the production of gene delivery particles cannot be delayed any longer. 1,12–16…”