Site-Specific Glycation and Chemo-enzymatic Antibody Sortagging for the Retargeting of rAAV6 to Inflamed Endothelium
Hannah A Pearce, Hongwei Qian, Timothy U Connell, Dexing Huang, Claudia Gottstein, Paul S Donnelly, Karlheinz Peter, Paul Gregorevic, Christoph E Hagemeyer
MOLECULAR THERAPY-METHODS & CLINICAL DEVELOPMENT | CELL PRESS | Published : 2019
Gene therapy holds great potential for conditions such as cardiovascular disease, including atherosclerosis and also vascular cancers, yet available vectors such as the adeno-associated virus (rAAV) transduce the vasculature poorly. To enable retargeting, a single-chain antibody (scFv) that binds to the vascular cell-adhesion molecule (VCAM-1) overexpressed at areas of endothelial inflammation was site specifically and covalently conjugated to the exterior of rAAV6. To achieve conjugation, the scFv was functionalized with an orthogonal click chemistry group. This conjugation utilized site-specific sortase A methodology, thus preserving scFv binding capacity to VCAM-1. The AAV6 was separately..View full abstract
This work was funded by the National Health and Medical Research Council (NHMRC), the Australian Research Council (ARC), and the National Heart Foundation of Australia (NHF). The work was also supported in part by the Victorian Government's Operational Infra-structure Support Program. We thank Prof. Kleinschmidt for the anti-AAV producing hybridoma cell lines.