2nd Annual DNA Process Development & Manufacturing Summit

Title: Optimization of Plasmid DNA (pDNA) Extraction via Lysis to Enhance Manufacturing Efficiency and Quality

Speakers:

Matevž Korenč, Product Manager for Alkalizator, Sartorius BIA Separations

Robin Pelizzari, Scientist pDNA Small Scale DSP Development, mRNA Center of Excellence, Sanofi Vaccine, France

Abstract:

Plasmid DNA (pDNA) isolation involves a crucial cell lysis step using a strong alkaline solution, followed by neutralization and precipitation to recover target pDNA while removing cellular contaminants like genomic DNA and proteins. Uncontrolled alkaline use and mechanical stress can degrade pDNA, and highly viscous solutions may cause heterogeneities or require extensive mixing, further compromising pDNA integrity.

Taking these aspects into consideration, we will present challenges and optimization opportunities in the context of scaling-up pDNA process for mRNA applications. Key considerations include maximizing product yield and quality while minimizing residual impurities, managing product/waste volumes and ensuring process step robustness.

As part of our evaluation of pDNA extraction methods, we will present an automated in-line lysis system (Alkalizator from Sartorius BIA Separations) with a closed single-use loop, tailored for process development up to pilot-scale within a GMP environment. It features advanced mixing control, enabling automatic, continuous processing of up to 500 liters of resuspended cells per workday, depending on configuration.

We will demonstrate the system’s application in pDNA process, showcasing in-line lysis results including impact on pDNA quality.

Key Learning Objectives:  

• Comprehend the challenges associated with alkaline lysis in pDNA extraction.
• Evaluate the advantages of the Sartorius in-line lysis system in comparison to traditional batch lysis methods.
• Explore at-line in-process control strategies tailored for managing complex crude lysis samples.

About the event

The Inaugural DNA Process Development and Manufacturing Summit focused on boosting plasmid and cell-free DNA production while cutting costs and ensuring compliance. Over 3 days, attendees engaged with 100+ DNA production experts through workshops, presentations, and networking sessions, addressing key production challenges with insights from 18+ speakers from top companies like Sanofi, Roche, Genentech, and Pfizer.

Location: Boston, MA