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 withhigh selectivity and throughput. mRNA-based pharmaceuticals have potential to address a wide variety ofpathologies. mRNA-based vaccines can increase safety and dramatically shorten development timelines inpandemic scenarios. A number of mRNA-based COVID-19 vaccines are under development, and one suchvaccine has completed its Phase-I clinical trial and showed great promise as a response to the COVID-19pandemic. However, a company pioneering mRNA medicines has revealed that the lack of high throughputdownstream purification processes is a major hurdle that must be addressed in the upscaling of industrialmRNA production to yield the necessary quantity and quality. Considering the profound impact that COVID-19will have on the global population of nearly seven billion people, the time to develop a high productivity mRNApurification technology, like the one proposed, is now. By addressing this challenge, the proposed technologywill have a significant impact on mRNA production and, by association, improve patient accessibility to thevaccine. 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 viahybridization between adenine in the poly-A tail and deoxythymidine in oligo-dT. The goal of this CompetitiveRevision project is to demonstrate the feasibility of developing dT-based affinity membrane products with highbinding capacity for the rapid and selective purification of polyadenylated mRNA. Preliminary data are highlyencouraging. The products derived from this innovation will be first-in-market, disposable membranechromatography columns that can improve the mRNA purification productivity up to one hundred times withhigh 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 forcapture step purification of polyadenylated mRNA. In Specific Aim 1, Purilogics will evaluate the roles playedby ligand structure and density, synthesis conditions, and bind-and-elute conditions on capacity and recoveryusing a commercially available purified mRNA. In Specific Aim 2, Purilogics will collaborate with a partnercontract manufacturing organization to quantify membrane column performance for capture step purification ofpolyadenylated mRNA prepared with in vitro transcription (IVT) processes. The prototypes also will bebenchmarked against existing products. Multiple iterations of synthesis and performance characterization willimprove membrane performance. Immediate market entry for the new column products will be sales topurification scientists and engineers in biopharmaceutical companies.