A Novel Automated Alkaline Lysis Solution For cGMP Manufacturing

Nucleic acid therapeutics, encompassing plasmids and mRNA, represent a promising new frontier in drug development. The success of mRNA vaccines against SARS-CoV-2 exemplifies this potential. However, large-scale production of these drugs faces challenges, particularly in plasmid purification. The current lysis step, often reliant on manual intervention, hinders process consistency and speed. To address this, we propose a fully automated alkaline lysis solution. This system offers significant advantages in terms of speed, batch-to-batch consistency, and scalability.

Through optimization of key parameters, such as the P1 buffer volume per gram of wet cell paste (WCP) and sodium hydroxide (NaOH) concentration in P2 buffer, we achieved a plasmid recovery rate exceeding 40%. Furthermore, the automated system ensures consistent lysis efficiency exceeding 90% across batches. This allows for processing of large-scale fermentations (50-100L) and production of 4-6 grams of supercoiled plasmid per batch, with host cell DNA (HCD) impurities below 0.2%. Discover how this automated approach offers a significant improvement for efficient and cost-effective plasmid purification, paving the way for broader adoption of nucleic acid-based therapeutics.