Adeno-Associated Virus 5 Protein Particles Produced by <i>E. coli</i> Cell-Free Protein Synthesis.

ABSTRACT: Recombinant adeno-associated viruses (rAAVs) have emerged as important tools for gene therapy and, more recently, vaccine development. Nonetheless, manufacturing can be costly and time-consuming, emphasizing the importance of alternative production platforms. We investigate the potential of E. coli-based cell-free protein synthesis (CFPS) to produce recombinant AAV5 virus-like particles (VLPs). AAV5 virus protein 3 (VP3) constructs, both with and without Strep-tag II, were expressed with CFPS. Lower reaction temperatures resulted in increased solubility, with the untagged variant containing nearly 90% more soluble VLP VP3 protein at 18 °C than at 37 °C. Affinity chromatography of N-terminally Strep(II)-tagged VP3 enabled successful isolation with minimal processing. DLS and TEM conf

SUBMITTER: Deuker D 

PROVIDER: S-EPMC11421080 | biostudies-literature | 2024 Sep

REPOSITORIES: biostudies-literature