Small synthetic ligands for the enrichment of viral particles pseudotyped with amphotropic murine leukemia virus envelope

Abstract

Retroviral vectors gained popularity toward other viral vectors as they integrate their genome into hosts’ genome, a characteristic required for the modification of stem cells. However, the production of viable particles for gene therapy is hampered by the low ratio of infectious to non-infectious viral particles after purification, low titers and limited number of competent viral receptors. We have developed de novo two fully synthetic triazine-based ligands that can selectively bind retroviral particles pseudotyped with amphotropic murine leukemia virus envelope (AMPHO4070A). A 78-membered library of triazine-based ligands was designed in silico and was virtually screened against the modeled structure of the AMPHO4070A protein. Ligands displaying the highest energy of binding were synthesized on cross-linked agarose and experimentally tested. Adsorbents containing ligands A5A10 and A10A11 showed selectivity toward viral particles containing the target protein (VLP-AMPHO), binding 19±5μg/g support and 47±13μg/g support, respectively. The elution conditions for both ligands were mild and with high recovery yields (80–100%), in comparison with common purification practices. These results were based on a lab-scale experimental setting with VLP integrity being confirmed through TEM. In particular, the elution buffer containing 12mM imidazole allowed the recovery of intact amphotropic viral particles.