title: A novel anti-VEGF165 monoclonal antibody-conjugated liposomal nanocarrier system: physical characterization and cellular uptake evaluation in vitro and in vivo.
abstract: Vascular endothelial growth factor (VEGF) is an important target for cancer therapy. In the present study, we conjugated the novel fully-human anti-VEGF165 monoclonal antibody, mAb165, with a PEGylated liposome (lip) to produce a monoclonal antibody-conjugated PEGylated liposome (mAb-lip). Physical characterization of mAb-lips showed an average particle size of 108nm. Using a bicinchoninic acid (BCA) assay, the coupling efficiency of mAb165 conjugated to the liposome was 69.8±0.5μg mAb/μmol phospholipid. In addition, we confirmed that conjugation between mAb165 and the liposome did not affect the structure and VEGF binding affinity of the antibody. Cell uptake of mAb-lips was assessed in four cell lines: MCF-7, HepG-2, SGC-7901, and L02 cells. Confocal microscopy and flow cytometry demonstrated that there was no significant difference in cell uptake between mAb-lips and mAb-free liposome either in VEGF-expressing tumor cells or normal cells. Moreover, the cytotoxicity of paclitaxel encapsulated in mAb-lips was not increased in the four cell lines. However, in BALB/c nude mice bearing MCF-7 xenografts, mAb-lips showed superior targeting activity to tumor tissues when compared with the unmodified liposome, which was demonstrated by the fact that rhodamine-labeled mAb-lips exhibited higher fluorescence intensity in tumor tissues than the unmodified liposome. Thus, our study indicated that mAb-lips may have the potential to enhance the therapeutic index of anticancer agents through targeted delivery to tumor cells in vivo.
authors: Department of Biochemistry, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, PR China.
authors: Chenyang
authors: C
authors: Shi
authors: Department of Biochemistry, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, PR China.
authors: Fei
authors: F
authors: Gao
authors: Department of Biochemistry, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, PR China. Electronic address: xdgao@cpu.edu.cn.
authors: Xiangdong
authors: X
authors: Gao
authors: Department of Biochemistry, School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, PR China. Electronic address: liuyuyaoda@163.com.
authors: Yu
authors: Y
authors: Liu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D016923:Cell Death / Q000187:drug effects; D004337:Drug Carriers / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis / Q000187:drug effects*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D056945:Hep G2 Cells; D006801:Humans; D008081:Liposomes / Q000737:chemistry* / Q000648:ultrastructure; D061986:MCF-7 Cells; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D042461:Vascular Endothelial Growth Factor A / Q000276:immunology* / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
chemical_list: D000911:Antibodies, Monoclonal; D004337:Drug Carriers; D008081:Liposomes; D042461:Vascular Endothelial Growth Factor A
keywords: Cellular uptake; Cytotoxicity; PEGylated liposomes; Targeted delivery; VEGF mAb