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.
Rüger
Ronny
R
Friedrich-Schiller-Universität Jena, Lehrstuhl für Pharmazeutische Technologie, Lessingstrasse 8, 07743 Jena, Germany.
Müller
Dafne
D
Fahr
Alfred
A
Kontermann
Roland E
RE
D016503:Drug Delivery Systems; D006801:Humans; D007128:Immunoglobulin Fragments / Q000008:administration & dosage* / Q000737:chemistry / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000008:administration & dosage* / Q000737:chemistry / Q000378:metabolism; D066298:In Vitro Techniques; D008081:Liposomes; D009571:Nitrilotriacetic Acid / Q000031:analogs & derivatives* / Q000737:chemistry; D009942:Organometallic Compounds / Q000737:chemistry*; D010316:Particle Size; D011485:Protein Binding; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism; D014280:Triglycerides / Q000737:chemistry*
D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D008081:Liposomes; D009942:Organometallic Compounds; D011994:Recombinant Proteins; D014280:Triglycerides; C055966:immunoglobulin Fv; C088169:1,2-dioleoyl-3-succinylglycerol; C088321:nickel nitrilotriacetic acid; D009571:Nitrilotriacetic Acid