Targeting of epidermal growth factor receptor (EGFR)-expressing tumor cells with sterically stabilized affibody liposomes (SAL).
Affibody molecules are small and stable antigen-binding molecules derived from the B domain of protein A. We applied a bivalent, high-affinity epidermal growth factor receptor (EGFR)-specific affibody molecule for the generation of targeted PEGylated liposomes. These sterically stabilized affibody liposomes (SAL) were produced by chemical coupling of the cysteine-modified affibody molecule to maleimide-PEG(2000)-DSPE and subsequent insertion into PEGylated liposomes. These SAL showed strong and selective binding to EGFR-expressing tumor cell lines. Binding was dependent on the amount of inserted affibody molecule-lipid conjugates and could be blocked by soluble EGF. Approximately 30% of binding activity was still retained after 6 days of incubation in human plasma at 37 degrees C. Binding of SAL to cells led to efficient internalization of the liposomes. Using mitoxantrone-loaded liposomes, we observed for SAL, compared to untargeted liposomes, an enhanced cytotoxicity toward EGFR-expressing cells. In summary, we show that SAL can be easily prepared from affibody molecules and thus may be suitable for the development of carrier systems for targeted delivery of drugs.
Institut fur Zellbiologie und Immunologie, Universitat Stuttgart, Allmandring 31, 70569 Stuttgart, Germany.
Julia
J
Beuttler
Miriam
M
Rothdiener
Dafne
D
Müller
Fredrik Y
FY
Frejd
Roland E
RE
Kontermann
D000906:Antibodies / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D001692:Biological Transport; D045744:Cell Line, Tumor; D003545:Cysteine / Q000737:chemistry; D066246:ErbB Receptors / Q000276:immunology* / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D020128:Inhibitory Concentration 50; D008081:Liposomes / Q000737:chemistry* / Q000378:metabolism; D008301:Maleimides / Q000737:chemistry; D008942:Mitoxantrone / Q000737:chemistry / Q000494:pharmacology; D049329:Nanostructures / Q000737:chemistry; D009369:Neoplasms / Q000473:pathology*; D010714:Phosphatidylethanolamines / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D013379:Substrate Specificity
D000906:Antibodies; D008081:Liposomes; D008301:Maleimides; D010714:Phosphatidylethanolamines; D011993:Recombinant Fusion Proteins; C038089:1,2-distearoylphosphatidylethanolamine; C043592:maleimide; D011092:Polyethylene Glycols; D008942:Mitoxantrone; D066246:ErbB Receptors; D003545:Cysteine