SBIR Phase II: Protein A Membrane Columns for Rapid Protein Purification

Project Summary This SBIR Urgent Competitive Revision will develop the first affinity membrane to purify therapeutic mRNA with high selectivity and throughput. mRNA-based pharmaceuticals have potential to address a wide variety of pathologies. mRNA-based vaccines can increase safety and dramatically shorten development timelines in pandemic scenarios. A number of mRNA-based COVID-19 vaccines are under development, and one such vaccine has completed its Phase-I clinical trial and showed great promise as a response to the COVID-19 pandemic. However, a company pioneering mRNA medicines has revealed that the lack of high throughput downstream purification processes is a major hurdle that must be addressed in the upscaling of industrial mRNA production to yield the necessary quantity and quality. Considering the profound impact that COVID-19 will have on the global population of nearly seven billion people, the time to develop a high productivity mRNA purification technology, like the one proposed, is now. By addressing this challenge, the proposed technology will have a significant impact on mRNA production and, by association, improve patient accessibility to the vaccine. Therapeutic mRNA usually possesses a polyadenylic acid (poly-A) tail. Oligo-deoxythymidine (oligo- dT) has been recognized as effective affinity ligand to isolate polyadenylated mRNA from feed streams via hybridization between adenine in the poly-A tail and deoxythymidine in oligo-dT. The goal of this Competitive Revision project is to demonstrate the feasibility of developing dT-based affinity membrane products with high binding capacity for the rapid and selective purification of polyadenylated mRNA. Preliminary data are highly encouraging. The products derived from this innovation will be first-in-market, disposable membrane chromatography columns that can improve the mRNA purification productivity up to one hundred times with high purity and yield compared to conventional resin columns. The Specific Aims of the study are to (1) synthesize and characterize mRNA affinity membranes and (2) test prototype affinity membrane columns for capture step purification of polyadenylated mRNA. In Specific Aim 1, Purilogics will evaluate the roles played by ligand structure and density, synthesis conditions, and bind-and-elute conditions on capacity and recovery using a commercially available purified mRNA. In Specific Aim 2, Purilogics will collaborate with a partner contract manufacturing organization to quantify membrane column performance for capture step purification of polyadenylated mRNA prepared with in vitro transcription (IVT) processes. The prototypes also will be benchmarked against existing products. Multiple iterations of synthesis and performance characterization will improve membrane performance. Immediate market entry for the new column products will be sales to purification scientists and engineers in biopharmaceutical companies.Public Health Relevance Statement mRNA-based vaccines are among the most promising solutions in response to today’s COVID-19 pandemic since they have the potential to increase vaccine safety and shorten vaccine timelines, leading to a more rapid and robust response to the pandemic. Unfortunately, current mRNA purification processes are time consuming and insufficient to produce pharmaceutical grade mRNA at industrial scale, which is needed for widespread vaccine distribution. The goal of this competitive revision is to develop an affinity membrane technology for rapid and selective purification of polyadenylated mRNA which will have a significant impact on expediting mRNA-based coronavirus vaccine production, and, by association, improving the general public’s accessibility to the vaccine.