Skip to main page content
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.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2013 Nov 18:103:2.17.1-2.17.57.
doi: 10.1002/0471142735.im0217s103.

Single-domain antibodies and their utility

Toya Nath Baral  1 , Roger MacKenzie  1   2 , Mehdi Arbabi Ghahroudi  1   2   3
Affiliations

Single-domain antibodies and their utility

Toya Nath Baral et al. Curr Protoc Immunol. .

Abstract

Engineered monoclonal antibody fragments have gained market attention due to their versatility and tailor-made potential and are now considered to be an important part of future immunobiotherapeutics. Single-domain antibodies (sdAbs), also known as nanobodies, are derived from VHHs [variable domains (V) of heavy-chain-only antibodies (HCAb)] of camelid heavy-chain antibodies. These nature-made sdAbs are well suited for various applications due to their favorable characteristics such as small size, ease of genetic manipulation, high affinity and solubility, overall stability, resistance to harsh conditions (e.g., low pH, high temperature), and low immunogenicity. Most importantly, sdAbs have the feature of penetrating into cavities and recognizing hidden epitopes normally inaccessible to conventional antibodies, mainly due to their protruding CDR3/H3 loops. In this unit, we will present and discuss comprehensive and step-by-step protocols routinely practiced in our laboratory for isolating sdAbs from immunized llamas (or other members of the Camelidae family) against target antigens using phage-display technology. Expression, purification, and characterization of the isolated sdAbs will then be described, followed by presentation of several examples of applications of sdAbs previously characterized in our laboratory and elsewhere.

Keywords: VHH; antibody fragment libraries dimeric VHH; heavy-chain antibodies; multivalency; pentabodies; single-domain antibody.

PubMed Disclaimer

References

Literature Cited

    1. Abulrob, A. , Sprong, H. , Van Bergen en Henegouwen, P. , and Stanimirovic, D. 2005. The blood-brain barrier transmigrating single domain antibody: Mechanisms of transport and antigenic epitopes in human brain endothelial cells. J. Neurochem. 95:1201-1214.
    1. Agrawal, V. , Slivac, I. , Perret, S. , Bisson, L. , St-Laurent, G. , Murad, Y. , Zhang, J. , and Durocher, Y. 2012. Stable expression of chimeric heavy chain antibodies in CHO cells. Methods Mol. Biol. 911:287-303.
    1. Aires da Silva, F. , Santa-Marta, M. , Freitas-Vieira, A. , Mascarenhas, P. , Barahona, I. , Moniz-Pereira, J. , Gabuzda, D. , and Goncalves, J. 2004. Camelized rabbit-derived VH single-domain intrabodies against Vif strongly neutralize HIV-1 infectivity. J. Mol. Biol. 340:525-542.
    1. Arbabi Ghahroudi, M. 1996. Generation and characterization of phage-displayed camel single-domain antibodies. Doctoral Dissertation, Vrije Universiteit Brussel, Brussels.
    1. Arbabi Ghahroudi, M. , Desmyter, A. , Wyns, L. , Hamers, R. , and Muyldermans, S. 1997. Selection and identification of single domain antibody fragments from camel heavy-chain antibodies. FEBS Lett. 414:521-526.

Substances

LinkOut - more resources