Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 115, Issue 9, Pages 2126-2131Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1722491115
Keywords
camelid antibody; horseradish peroxidase; GFP; nanobody; retina
Categories
Funding
- NIH [R37 NS029169]
- NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE [R37NS029169] Funding Source: NIH RePORTER
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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.
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