Skip Navigation
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Internet Explorer is no longer a supported browser.

This website may not display properly with Internet Explorer. For the best experience, please use a more recent browser such as the latest versions of Google Chrome, Microsoft Edge, and/or Mozilla Firefox. Thank you.

Your Environment. Your Health.

Publication Detail

Title: Generation, expression and utilization of single-domain antibodies for in vivo protein localization and manipulation in sea urchin embryos.

Authors: Schrankel, Catherine S; Gökirmak, Tufan; Lee, Chang-Wook; Chang, Geoffrey; Hamdoun, Amro

Published In Methods Cell Biol, (2019)

Abstract: Single-domain antibodies, also known as nanobodies, are small antigen-binding fragments (~15kDa) that are derived from heavy chain only antibodies present in camelids (VHH, from camels and llamas), and cartilaginous fishes (VNAR, from sharks). Nanobody V-like domains are useful alternatives to conventional antibodies due to their small size, and high solubility and stability across many applications. In addition, phage display, ribosome display, and mRNA/cDNA display methods can be used for the efficient generation and optimization of binders in vitro. The resulting nanobodies can be genetically encoded, tagged, and expressed in cells for in vivo localization and functional studies of target proteins. Collectively, these properties make nanobodies ideal for use within echinoderm embryos. This chapter describes the optimization and imaging of genetically encoded nanobodies in the sea urchin embryo. Examples of live-cell antigen tagging (LCAT) and the manipulation of green fluorescent protein (GFP) are shown. We discuss the potentially transformative applications of nanobody technology for probing membrane protein trafficking, cytoskeleton re-organization, receptor signaling events, and gene regulation during echinoderm development.

PubMed ID: 30948018 Exiting the NIEHS site

MeSH Terms: Animals; Camelids, New World/immunology; Camelus/immunology; Embryo, Nonmammalian/ultrastructure; Embryonic Development/genetics; Immunoglobulin Heavy Chains/genetics; Molecular Biology/methods*; Protein Transport/genetics; Protein Transport/immunology; Proteins/genetics; Proteins/isolation & purification*; Sea Urchins/growth & development; Sea Urchins/ultrastructure*; Single-Domain Antibodies/biosynthesis*; Single-Domain Antibodies/chemistry; Single-Domain Antibodies/immunology

Back
to Top