A Multi-attribute Method Supporting the Process Development for Non-viral Gene Therapy

Gene therapy has re-emerged as a new therapeutic modality due to the recent breakthroughs and the potential to provide a long-term treatment for a genetic disease for patients. Traditionally, viral vectors, such as adeno-associated virus (AAV), have been widely used but pose challenges such as potential humoral immune response, limitation of re-dose, and limited cargo capacity. In contrast, non-viral delivery with lipid nanoparticles (LNPs) can carry larger genetic payloads (> 10kb for DNA in theory) and the ability to re-dose the patients once the treatment wears off. In this work, we present a multi-attribute HPLC method which allows us to track the productivity of our nucleic acid product as well as provide a comprehensive impurity profile for in-process samples. The method can be implemented across the whole purification process with minimal sample manipulation and provides an overview of both the product of interest and process impurities within one assay. The method serves as a good in-process monitoring tool to ensure the batch-to-batch consistency of our process.