466. Automated Generation of Genetically Modified Human CD34+ Cells in a Functionally Closed System

Abstract

of different scales from 5 ml to 500 ml. Optimization of transfection parameters like cell density at the time of transfection, the ratio of helper to donor plasmid and total plasmid to PEI was conducted for each individual cell line. Lysates and supernatants were processed separately using gradient centrifugation and affinity chromatography. Purified virus stocks were then analyzed and titrated using methods to determine total viral particles, viral genomes and infectious titers. Preliminary results indicate that rAAV particles produced with suspension cells yield infectious titers equal or higher than particles produced by standard methods based on adherent cells. Further work is ongoing to analyze and implement this OSR platform. The presented process offers a novel method to produce rAAV for preclinical and clinical trials compliant to GMP, single-use and scalable.