Generation of immunoliposomes using recombinant single-chain Fv fragments bound to Ni-NTA-liposomes

Recombinant single-chain Fv antibody fragments (scFv) can be combined with liposomes to generate immunoliposomes for targeted drug delivery. Recent studies have shown that scFv molecules modified to express a C-terminal cysteine residue can be used for site-directed chemical conjugation. Here, we present a new method by immobilizing scFv fragments via their C-terminal hexahistidyl-tag on liposomes containing Ni-NTA-lipids (Ni-NTA-DOGS) in their lipid bilayer without the need to introduce additional reactive groups in the protein. Using an anti-endoglin scFv as a model antibody, we could show that scFv molecules are efficiently immobilized on the liposome surface and that these immunoliposomes bind specifically and strongly to endoglin-expressing endothelial cells. This approach allows for a rapid and flexible generation of target cell-specific immunoliposomes.