(1077-B) A Next-Generation Automated Platform for High-Throughput Commercial Production of Plasmid DNA

Abstract: The global demand for plasmid DNA has risen dramatically since the advent of cell and gene therapies (CGT) and has experienced hyper-growth following the outbreak of the COVID-19 pandemic. These cutting edge therapies come in many forms but all of them rely on plasmid DNA at some point either in their manufacture or mechanism of action. Unfortunately, scaling up manufacturing capacity for plasmid DNA is a difficult operation and it has become a bottleneck for the CGT industry. Contract development and manufacturing companies (CDMOs) can no longer meet the demand for high quality plasmid DNA due to substantial backlogs and the lack of automated solutions for plasmid manufacturing. This can significantly impede R&D pipelines and harm the expectations of the market, and most importantly that of the patients who are waiting for these therapies.
Viral vectors form the backbone of CGT and despite their considerable promise they are still beset by manufacturing challenges. The low yields of functional capsids containing the desired genetic payload remains a hurdle. Empty capsids require much larger doses to be delivered and this can provoke a strong immune response. New approaches in capsid engineering can yield millions of variant capsids which are screened for desirable properties. To systematically screen variants and scale up to 100mg-1g quantity of DNA for preclinical testing requires high-throughput plasmid manufacturing capabilities that almost all CGT companies presently lack. Here we present GenScript’s new automated plasmid production platform designed to produce up to 1000 ultra-low endotoxin plasmids at the mg scale per 24 hours. This is an integrated platform bringing the best of industrial automation to a biological manufacturing process. The platform consists of six industrial robot operated modules encompassing inoculation, incubation, lysis, chromatography and solvent extraction. It can operate continuously with in-process cleaning to prevent cross-contamination. We have demonstrated that we can obtain plasmid yields greater than 1mg with endotoxin levels < 0.1EU/μg and supercoiled content >90%. Our platform can produce plasmid libraries for capsid and vector screening, genome-wide CRISPR libraries, and in vitro transcription (IVT) templates for mRNA production for the CGT industry.