Biotinylated vectors as ligand screening platforms
Localization of adenovirus protein IX using biofilm associated protein and green fluorescent protein tagged capsomers and cryo-electron microscopy
Metabolic biotinylation allows one genetically engineered vector to be used with any biotinylated ligand. As such, biotinylated vectors can be used as a platform for combinatorial screening of both cell-targeting ligands and for receptors that are compatible with cell targeting. As proof of principle for this application, the team has screened for useful ligands to increase transduction of primary dendritic cells.
The following ligands were tested using the biotinylated fiber vector:
- Biotinylated antibodies against CD40, CD86, CD11b, CD11c and MHC II.
- Biotinylated protein ligands targeting toll-like receptors and scavenger receptors, such as heat shock 70kDa protein 8 (HSC70), aggregated albumin, biotinylated mannosylated bovine serum albumin targeting the mannose receptor, biotinylated asialofetuin targeting the dendritic asialofetuin receptor and biotinylated CpG oligonucleotides to target TLR9.
These diverse and structurally incompatible ligands were easily screened when complexed with biotinylated adenovirus using avidin as a bridge. Work is ongoing in the lab to use this technology to prescreen ligands on various targeting models.