Abstract
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.
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Cell Line, Tumor
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Drug Delivery Systems / methods*
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Endocytosis
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Flow Cytometry
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Humans
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Liposomes / immunology*
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Liposomes / isolation & purification
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Micelles
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Microscopy, Fluorescence
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Molecular Sequence Data
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Nanomedicine / methods
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Nanoparticles / chemistry*
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Neoplasms / metabolism*
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Organ Specificity
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Phosphatidylethanolamines / chemistry
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Plasmids / chemistry
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Plasmids / genetics
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Polyethylene Glycols / chemistry
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Receptors, Cell Surface / metabolism*
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Single-Chain Antibodies / metabolism*
Substances
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1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-methoxy-poly(ethylene glycol 2000)
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Liposomes
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Micelles
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Phosphatidylethanolamines
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Receptors, Cell Surface
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Single-Chain Antibodies
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Polyethylene Glycols