Antibody targeting of nanoparticles to tumor-specific receptors: immunoliposomes.
Immunoliposomes generated by coupling of antibodies to the liposomal surface allow for an active tissue targeting, e.g., through binding to tumor cell-specific receptors. Instead of whole antibodies, single-chain Fv fragments (scFv), which represent the smallest part of an antibody containing the entire antigen-binding site, find increasing usage as targeting moiety. Here we provide protocols for the preparation of type II scFv immunoliposomes by the conventional coupling method as well as the post-insertion method. Furthermore protocols to analyze binding of these immunoliposomes to antigen-expressing cells as well as internalization through receptor-mediated endocytosis are included.
Rothdiener
Miriam
M
Institute of Cell Biology and Immunology, University of Stuttgart, Stuttgart, Germany.
Beuttler
Julia
J
Messerschmidt
Sylvia K E
SK
Kontermann
Roland E
RE
D000595:Amino Acid Sequence; D001483:Base Sequence; D045744:Cell Line, Tumor; D016503:Drug Delivery Systems / Q000379:methods*; D004705:Endocytosis; D005434:Flow Cytometry; D006801:Humans; D008081:Liposomes / Q000276:immunology* / Q000302:isolation & purification; D008823:Micelles; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D050997:Nanomedicine / Q000379:methods; D053758:Nanoparticles / Q000737:chemistry*; D009369:Neoplasms / Q000378:metabolism*; D009928:Organ Specificity; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D011092:Polyethylene Glycols / Q000737:chemistry; D011956:Receptors, Cell Surface / Q000378:metabolism*; D057127:Single-Chain Antibodies / Q000378:metabolism*
C519184:1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-methoxy-poly(ethylene glycol 2000); D008081:Liposomes; D008823:Micelles; D010714:Phosphatidylethanolamines; D011956:Receptors, Cell Surface; D057127:Single-Chain Antibodies; D011092:Polyethylene Glycols