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Proc Natl Acad Sci U S A. 2018 Feb 27;115(9):2126-2131. doi: 10.1073/pnas.1722491115. Epub 2018 Feb 13.

Reporter-nanobody fusions (RANbodies) as versatile, small, sensitive immunohistochemical reagents.

Author information

1
Center for Brain Science, Harvard University, Cambridge MA, 02138.
2
Department of Molecular and Cellular Biology, Harvard University, Cambridge MA, 02138.
3
Center for Brain Science, Harvard University, Cambridge MA, 02138; sanesj@mcb.harvard.edu.

Abstract

Sensitive and specific antibodies are essential for detecting molecules in cells and tissues. However, currently used polyclonal and monoclonal antibodies are often less specific than desired, difficult to produce, and available in limited quantities. A promising recent approach to circumvent these limitations is to employ chemically defined antigen-combining domains called "nanobodies," derived from single-chain camelid antibodies. Here, we used nanobodies to prepare sensitive unimolecular detection reagents by genetically fusing cDNAs encoding nanobodies to enzymatic or antigenic reporters. We call these fusions between a reporter and a nanobody "RANbodies." They can be used to localize epitopes and to amplify signals from fluorescent proteins. They can be generated and purified simply and in unlimited amounts and can be preserved safely and inexpensively in the form of DNA or digital sequence.

KEYWORDS:

GFP; camelid antibody; horseradish peroxidase; nanobody; retina

PMID:
29440485
PMCID:
PMC5834735
DOI:
10.1073/pnas.1722491115
[Indexed for MEDLINE]
Free PMC Article

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