Fast-Track Innovation for Plasmid DNA Production

The Sartorius team in Slovenia developed a fully automated system for bacterial cell lysis in plasmid DNA production. Read how they are part of the solution in producing new therapies by addressing a pain point in downstream processing.

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Plasmid DNA (pDNA) has a wide range of applications: It is used as a vaccine, as a transfection vector for the preparation of viral vectors for gene therapies (e.g. AAV viruses) and as starting material for the preparation of mRNA vaccines.

It is manufactured by genetically engineering bacteria to replicate a desired plasmid. An important step in this is the so-called alkaline cell lysis: After cell harvest, bacterial cells are exposed to a strong alkaline solution, which breaks up cell walls to release pDNA. To do this, the cells are traditionally broken down (lysed) in batches. Sounds complex? It is complex.

Identifying and Filling a Gap

"Alkaline cell lysis during plasmid production is a technologically challenging process, especially when working with large volumes," explains Dr. Matevž Korenč, Product Manager at Sartorius. "It is important to keep precise control over pH and temperature during exposure to strong base and control the neutralization process. Efficient and low-shear mixing is crucial to reduce potential damage to the target plasmids."

The challenge: Keeping precise control over pH and temperature during exposure to strong base, and control the neutralization process.

Already since the late 1990s, the team in Slovenia has been working on pDNA process development. What initially started with chromatography purification, has been expanded to cover all downstream purification aspects. Today, with Cornerstone^® Biomanufacturing Development Services, the team of Sartorius BIA Separations accelerates process development from pre-clinical to market supply. 

With the recent uptake and internalization of pDNA production by gene therapy and mRNA developers, the team in Slovenia supports many of them with the development of pDNA processes and identified a significant technological gap in this context – the market didn't offer a solution for reliable and scalable alkaline lysis. So, the team in Slovenia started to develop a solution themselves to fill this gap – and the Alkalizator was born: a fully automated system for dissolving and removing host cells during pDNA production.

This optimization directly reduces the production costs, making the development of therapies for rare genetic diseases more attractive to biopharma companies – and treatments ultimately more affordable for patients.

Dr. Matevž Korenč, Product Manager  at Sartorius BIA Separations

"The system gives users precise control over the alkaline process, resulting in higher efficiency and quality of the pDNA compared to the traditional batch-lyses approach," explains Matevž. "This optimization directly reduces the production costs, making the development of therapies for rare genetic diseases more attractive to biopharma companies – and treatments ultimately more affordable for patients."

From E. coli to Purified Plasmid

pDNA molecules are larger and more sensitive than traditional modalities, creating a new set of manufacturing challenges. Explore the steps in a typical pDNA production process.

Learn More

Fast Results Through Close Collaboration

Typically, when developing new products, the journey from idea to a market-ready product spans years. Recurring consultations and feedback rounds are major time-consuming factors. In case of the Alkalizator, development started in late 2021 and by December 2023, the first GMP-compliant system had been put into action, with more units undergoing testing in North America and Europe. After the pre-launch of the system at a scientific symposium in June 2024, four units were already used in process development and three units in GMP-production by the end of the year.

"Having experts and customers as developing partners working towards the same goal under the same roof really helped to speed up the process significantly," Matevž says. Also, people from other R&D teams at Sartorius were involved: The Alkalizator is for example equipped with Sartorius flow sensors, which are critical for high accuracy of lysis procedure.
 

We're a very young and well-trained team here in Slovenia and have a “Moving-Things-Fast-Attitude.

Dr. Matevž Korenč, Product Manager  at Sartorius BIA Separations

Not to forget for fast development: An ambitious team. "We’re a very young and well trained team here in Slovenia and have a "Moving-Things-Fast"-Attitude. Working with cutting-edge technologies and being surrounded by so many smart people that work together on improving the health of people is something I really enjoy about working here," Matevž says.

And the work continues: "The current product is a compact and modular instrument that can easily be upgraded. We are already working on scaled-down and scaled-up units and an integration with in-line filtration," he adds.