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    Biotinylated Bionanocapsules for Displaying Diverse Ligands Toward Cell-specific Delivery
    The Journal of Biochemistry (2009)
    Takuya ShishidoY. AzumiTakeshi NakanishiMitsuo UmetsuTsutomu Tanaka
    Bionanocapsule (BNC) is hollow nanoparticle composed of the l-protein of the hepatitis B virus surface antigen. BNC allows targeted delivery of either genes or drugs only to hepatocytes, but not to other cell types. In this study, we attempted to alter the specificity of BNC by insertion of biotin-acceptor peptide (BAP), which is efficiently biotinylated using biotin ligase BirA from Escherichia coli. Using streptavidin as a linker, biotinylated BNC could be display various biotinylated ligands that are otherwise difficult to fuse with BNC, such as antibodies, synthetic peptides and functional molecules. BAP-fused BNC was efficiently biotinylated and effectively displayed streptavidin. Furthermore, we demonstrated that biotinylated BNC was internalized into targeted cells via biotinylated Nanobody displayed on the BNC surface. Biotinylated BNC permit display of diverse ligands, and thus have potential as a versatile carrier for drug delivery to a variety of target cells.
    Streptavidin
    Linker
    Citations (15)
    Antibodies to biotin enable large-scale detection of biotinylation sites on proteins
    Nature Methods (2017)
    Namrata D. UdeshiKayvon PedramTanya SvinkinaShaunt FereshetianSamuel A. Myers
    Streptavidin
    Citations (136)
    [The electrophoresis of biotinylated nucleic acids].
    PubMed (1992)
    Popa LlAdriana PleșaR RepanoviciPopa Lm
    A simple and rapid method was worked out to evaluate the biotinylation level of the pBR322 and pSVK1H genetic cloning vectors, using gel electrophoresis. Avidin was used to slow down the migration of biotinylated DNA: the DNA migration speed diminished as the biotinylation level rose, due to DNA complexation. The highest level of biotinylation is characterized by the formation of a biotinylated nucleic acid-avidin complex with no electrophoretical mobility.
    Avidin
    Cloning (programming)
    Citations (0)
    In Vivo Biotinylated scFv Fragments
    Springer eBooks (2010)
    Laila Al-HalabiTorsten Meyer
    Streptavidin
    Avidin
    Protein Engineering
    Target protein
    Citations (1)
    Short-term culture of surface-biotinylated cells: Application in non-radioactive analysis of surface protein shedding
    Immunology Letters (1995)
    Stefan HausmannRenate ClausHermann Walzel
    Immunoprecipitation
    Streptavidin
    Citations (6)
    Biotinylation of Proteins in Solution and on Cell Surfaces
    Current Protocols in Protein Science (1996)
    Elizabeth J. Luna
    This unit contains three protocols for biotinylation of isolated proteins: attaching biotin to primary amines (e.g., amino groups of lysyl residues); attaching biotin to sulfhydryls (i.e., thiol groups of cysteinyl residues); and attaching biotin to carbohydrate residues on proteins. As biotinylation of lysyl and cysteinyl residues may alter protein function, modification of protein carbohydrates, which is usually innocuous, may be preferable if intact protein function is required (e.g., for activity assays or affinity purification). Biotinylation of cysteinyl thiols requires that disulfide bonds in isolated proteins be reduced before labeling. Biotinylation of surface proteins on living cells is also described using mild reaction conditions. Finally, this unit includes a brief description of methods for detecting biotinylated proteins.
    Citations (10)
    Biotinylated erythrocytes: in vivo survival and in vitro recovery
    Blood (1987)
    T SuzukiGL Dale
    Avidin
    Citations (6)
    Position-specific incorporation of biotinylated non-natural amino acids into a protein in a cell-free translation system
    Biochemical and Biophysical Research Communications (2007)
    Takayoshi WatanabeNorihito MuranakaIssei IijimaTakahiro Hohsaka
    Citations (21)
    Biotinylated erythrocytes: in vivo survival and in vitro recovery
    Blood (1987)
    T SuzukiGL Dale
    Avidin
    Citations (112)
    Use of Biotinylated Chitosan for Substrate-Mediated Gene Delivery
    Bioconjugate Chemistry (2012)
    Wei‐Wen HuWun-Jheng SyuWen‐Yih ChenRuoh‐Chyu RuaanYu‐Che Cheng
    To improve transfection efficiency of nonviral vectors, biotinylated chitosan was applied to complex with DNA in different N/P ratios. The morphologies and the sizes of formed nanoparticles were suitable for cell uptake. The biotinylation decreased the surface charges of nanoparticles and hence reduced the cytotoxicity. The loading capacities of chitosan were slightly decreased with the increase of biotinylation, but most of the DNA molecules were still complexed. Using different avidin-coated surfaces, the interaction between biotinylated nanoparticles to the substrate may be manipulated. The in vitro transfection results demonstrated that biotinylated nanoparticles may be bound to avidin coated surfaces, and the transfection efficiencies were thus increased. Through regulating the N/P ratio, biotinylation levels, and surface avidin, the gene delivery can be optimized. Compared to the nonmodified chitosan, biotinylated nanoparticles on biomaterial surfaces can increase their chances to contact adhered cells. This spatially controlled gene delivery improved the gene transfer efficiency of nonviral vectors and could be broadly applied to different biomaterial scaffolds for tissue engineering applications.
    Avidin
    Biomaterial
    Citations (27)