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

Citation:
Fernandes, CSM, Castro R, Coroadinha AS, Roque ACA.  2016.  Small synthetic ligands for the enrichment of viral particles pseudotyped with amphotropic murine leukemia virus envelope. Journal of Chromatography A. 1438:160–170.: Elsevier B.V.

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 ± 5 $μ$g/g support and 47 ± 13 $μ$g/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 12 mM imidazole allowed the recovery of intact amphotropic viral particles.

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