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  • Open Access
    Authors: 
    Jie Xian Dong; Yongam Lee; Michael Kirmiz; Stephanie Palacio; Camelia Dumitras; Claudia M. Moreno; Richard Sando; L. Fernando Santana; Thomas C. Südhof; Belvin Gong; +2 more
    Publisher: eScholarship, University of California
    Country: United States
    Project: NIH | Recombinant Immunolabels ... (1U24NS109113-01), NIH | Administrative Supplement... (3U01NS099714-02S1)

    Nanobodies (nAbs) are small, minimal antibodies that have distinct attributes that make them uniquely suited for certain biomedical research, diagnostic and therapeutic applications. Prominent uses include as intracellular antibodies or intrabodies to bind and deliver cargo to specific proteins and/or subcellular sites within cells, and as nanoscale immunolabels for enhanced tissue penetration and improved spatial imaging resolution. Here, we report the generation and validation of nAbs against a set of proteins prominently expressed at specific subcellular sites in mammalian brain neurons. We describe a novel hierarchical validation pipeline to systematically evaluate nAbs isolated by phage display for effective and specific use as intrabodies and immunolabels in mammalian cells including brain neurons. These nAbs form part of a robust toolbox for targeting proteins with distinct and highly spatially-restricted subcellular localization in mammalian brain neurons, allowing for visualization and/or modulation of structure and function at those sites.

Include:
1 Research products, page 1 of 1
  • Open Access
    Authors: 
    Jie Xian Dong; Yongam Lee; Michael Kirmiz; Stephanie Palacio; Camelia Dumitras; Claudia M. Moreno; Richard Sando; L. Fernando Santana; Thomas C. Südhof; Belvin Gong; +2 more
    Publisher: eScholarship, University of California
    Country: United States
    Project: NIH | Recombinant Immunolabels ... (1U24NS109113-01), NIH | Administrative Supplement... (3U01NS099714-02S1)

    Nanobodies (nAbs) are small, minimal antibodies that have distinct attributes that make them uniquely suited for certain biomedical research, diagnostic and therapeutic applications. Prominent uses include as intracellular antibodies or intrabodies to bind and deliver cargo to specific proteins and/or subcellular sites within cells, and as nanoscale immunolabels for enhanced tissue penetration and improved spatial imaging resolution. Here, we report the generation and validation of nAbs against a set of proteins prominently expressed at specific subcellular sites in mammalian brain neurons. We describe a novel hierarchical validation pipeline to systematically evaluate nAbs isolated by phage display for effective and specific use as intrabodies and immunolabels in mammalian cells including brain neurons. These nAbs form part of a robust toolbox for targeting proteins with distinct and highly spatially-restricted subcellular localization in mammalian brain neurons, allowing for visualization and/or modulation of structure and function at those sites.

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