A toolbox of nanobodies developed and validated for use as intrabodies and nanoscale immunolabels in mammalian brain neurons

Jie Xian Dong, Yongam Lee, Michael Kirmiz, Stephanie Palacio, Camelia Dumitras, Claudia M. Moreno, Richard Sando, L. Fernando Santana, Thomas C. Südhof, Belvin Gong, Karl D. Murray, James S. Trimmer

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

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.

Original languageEnglish (US)
Article numbere48750
JournaleLife
Volume8
DOIs
StatePublished - Sep 2019

ASJC Scopus subject areas

  • Neuroscience(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

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    Dong, J. X., Lee, Y., Kirmiz, M., Palacio, S., Dumitras, C., Moreno, C. M., Sando, R., Santana, L. F., Südhof, T. C., Gong, B., Murray, K. D., & Trimmer, J. S. (2019). A toolbox of nanobodies developed and validated for use as intrabodies and nanoscale immunolabels in mammalian brain neurons. eLife, 8, [e48750]. https://doi.org/10.7554/eLife.48750