From early development stage until large-scale GMP plasmid production
In the pharmaceutical industry, plasmid DNA (abbreviated as pDNA) serves various crucial purposes in the manufacturing of biopharmaceuticals: pDNA is at the heart of DNA vaccines and functions as critical starting material for Cell and Gene Therapy (e.g., the production of viral vectors or in-vitro transcribed mRNA).
Eurofins CDMO has the required expertise to support pDNA projects from early development stage until large-scale GMP plasmid production.
Strain and clone selection
Strain and clone selection are critical steps in the pDNA manufacturing process as they can substantially influence the yield, quality and scalability of the final product.
Within the Eurofins network, custom plasmids can be tailored to the specific requirements of a project. This includes selecting appropriate promotors, antibiotic resistance markers, codon optimization, etc.
During strain selection, the choice of the host organism will be investigated to obtain the best results in terms of genetic stability, and the ability to support high-density cultures to enhance productivity.
Following the strain selection, a screening is performed to identify clones exhibiting the best plasmid stability, productivity and absence of undesired mutations. Upon selection of the best clone, a Research Cell Bank (RCB) will be produced. According to the specific project requirements, such RCB can also be the starting point for further banking activities, up to the manufacturing of a fully characterized, GMP grade, Master and Working Cell Bank (MCB, WCB).
Process & analytical development
In the process development phase, the aim is to create a robust, scalable, and economically viable process to deliver high quality plasmid DNA meeting to regulatory standards.
Using a parallel bioreactor system, fermentation conditions including growth media, temperature, pH, aeration and agitation can be optimized to maximize cell growth and pDNA yield. The downstream processing part will involve the purification and isolation of plasmid DNA from the cellular components and contaminants produced during the upstream fermentation process to meet the set quality requirements.
At every stage, critical process parameters will be identified and optimized to find an optimal balance between quality and yield. To this end, analytical methods will be employed to continuously monitor quality attributes during manufacturing.
Research-grade production
As an economical alternative to GMP grade manufacturing, we developed an efficient plasmid DNA manufacturing platform that can be used to produce plasmid DNA within short timelines in a research-grade setting. The process is designed to meet the QbD principles and will allow a seamless transition of the project to GMP. The process entails:
- High cell density fermentation (up to 100L in stainless steel fermenter)
- Scalable alkaline lysis followed by an optimized clarification method resulting in highly purified supercoiled plasmid DNA, and low genomic DNA and RNA content
- Chromatography step(s) for contaminant removal
- Final 0.2 µm filtration and filling of the bulk solution
Appropriate analytics focused on safety, identity, and purity testing, as well as stability testing will be conducted in a research-grade environment.
Deliverables:
- Batch record documents
- Manufacturing report
- Certificate of Testing
GMP Manufacturing
Either starting from the materials provided by our customers, or starting from the materials generated with the previously describer development activities, Eurofins CDMO offers a comprehensive suite of services tailored for the production of plasmid DNA under strict GMP regulations.
- E.coli cell banking services
- for master (MCB) and working (WCB) cell banks
- Storage in temperature-controlled GMP warehouse
- GMP grade plasmid production
- Using complete single use process from USP until DSP
- From mg to g scale
- Qualified equipment in Grade C environment
- Quality control for MCB, in-process testing, plasmid release and characterization
- Incoming QC of raw materials
- Assay qualification
- Stability testing
- QP release and full documentation
- QA reviewed batch record documents
- Certificate of conformity
- TSE/BSE certificate
- Certificate of analysis
Integrated solutions for your plasmid DNA
Eurofins can offer an integrated solution to its customers from the development and manufacturing of plasmid DNA, including sterile Fill & finish as well as clinical supply services.
To learn more