This paid-for advertorial by Catalent appeared in DDW Volume 24 – Issue 1, Winter 2022/2023
One significant challenge in the manufacturing of cell and gene therapies is the production of high-quality plasmid DNA (pDNA). pDNA is a critical raw material for advanced therapeutics including mRNA and viral vector-based cell and gene therapies and vaccines.
The market for cell and gene therapies has grown rapidly over the last few years and will likely continue, with forecasts predicting it will grow 39% to around $42.56 billion by 2030 (BioSpace, 11 August 2022). This rapid emergence of cell and gene therapies has fuelled demand for plasmids and has created a manufacturing ‘bottleneck’.
Plasmid DNA: a critical reagent
pDNA is the starting material for the development of many cell and gene therapy products, irrespective of the mode of action. Figure 1 shows the development pathways for non-viral and viral vector gene therapies, and cell therapy and illustrates the critical role pDNA plays in their development.
At its 17,000-square-foot facility, on the cell therapy campus in Gosselies, Belgium, Catalent provides end-to-end capabilities that span from preclinical up to commercial supply, using fully single-use technology.
Plasmid DNA platform process
The pDNA platform process includes cell bank creation, upstream and downstream activities, and analytical testing.
The starting point of the upstream process is the development of the E. coli cell bank that contains the plasmid. E. coli cells are first grown in shake flasks to produce the necessary biomass required to inoculate a fermenter. The harvested cells are lysed through a controlled alkaline treatment, which is key to recover as much product as possible without degrading the plasmid molecules. A clarification step to remove cell debris is needed before commencing the downstream process.
The first step in the downstream process is to concentrate the lysate to a more practical volume and buffer exchange in a solution that is appropriate for chromatography. This chromatographic step is used to clear any remaining E. coli residual contaminants as well as product-related impurities. The plasmids are then formulated into the desired formulation buffer through an ultrafiltration/diafiltration (UF/DF) step. A 0.2 µm filtration step completes the process. Samples are then sent for analytical testing.
Fully integrated pDNA services
Catalent uses a flexible approach to support innovators which can begin at the lead identification step (including the development of E. coli cell lines), or later in the process using existing GMP cell banks for programs that have already completed early phase trials, for example. Once leads are identified, Catalent’s cell line, process, and quality control assay development services will help ensure the plasmids meet the target critical quality attributes.
Additionally, Catalent can support development and manufacturing activities across the entire cell and gene therapy product spectrum from pDNA, mRNA, and viral vectors, to autologous and allogeneic cell therapies. Drug developers can choose to work with Catalent on one or two of these modalities, at varying stages of the process, or benefit from the integration of development and production services across product modalities, which can lead to shortened project timelines.
Find out more
Matthias Craig, Senior Manager, Product Development, Catalent Cell & Gene Therapy recently discussed the pDNA platform process in a webinar available to watch on-demand.