Abstract
The preparation of antibody libraries starting from lymphocytes recovered from immunized members of the Camelidae enables to collect binders that underwent somatic maturation. However, the time and costs necessary to prepare a library for each new antigen may urge to look for alternatives such as those offered by large one-pot libraries. Here we describe how to obtain a suitable naïve library using material from nonimmunized llamas. Despite the lack of somatic maturation, the selection based on phage display allowed to isolate from such naïve libraries VHHs with affinity in the subnanomolar range and suitable for the standard immunoapplications.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Arbabi Ghahroudi M et al (1997) Selection and identification of single domain antibody fragments from camel heavy-chain antibodies. FEBS Lett 414:521–526
Muyldermans S (2001) Single domain camel antibodies: current status. J Biotechnol 74:277–302
Yeo C, Saunders N, Locca D (2009) Ficoll-Paque™ versus Lymphoprep™: A comparative study of two density gradient media for therapeutic bone marrow mononuclear cell preparations. Regen Med 4:689–696
Olichon A, Surrey T (2007) Selection of genetically encoded fluorescent single domain antibodies engineered for efficient expression in Escherichia coli. J Biol Chem 282:36314–36320
Garaicoechea L et al (2008) Llama-derived single-chain antibody fragments directed to rotavirus VP6 protein possess broad neutralizing activity in vitro and confer protection against diarrhea in mice. J Virol 82:753–764
Bossi S et al (2010) Antibody-mediated purification of co-expressed antigen-antibody complexes. Protein Expr Purif 72:55–58
Aliprandi M et al (2010) The availability of a recombinant anti-SNAP antibody in VHH format amplifies the application flexibility of SNAP-tagged proteins. J Biomed Biotechnol. doi:10.1155/2010/658954
Monegal A et al (2009) Immunological applications of single-domain llama recombinant antibodies isolated from a naive library. Prot Engineer Des Sel 22:273–280
Nguyen VK, Muyldermans S, Hamers R (1998) The specific variable domain of camel heavy-chain antibodies is encoded in the germline. J Mol Biol 275:413–418
Skerra A, Plückthun A (1988) Assembly of a functional immunoglobulin Fv fragment in Escherichia coli. Science 240:1038–1041
Dong J et al (2010) A single-domain llama antibody potently inhibits the enzymatic activity of Botulinum neurotoxin by binding to the non-catalytic α–exosite binding region. J Mol Biol 397:1106–1118
Deschacht N et al (2010) A novel promiscuous class of camelid single-domain antibody contributes to the antigen-binding repertoire. J Immunol 184:5696–5704
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Olichon, A., de Marco, A. (2012). Preparation of a Naïve Library of Camelid Single Domain Antibodies. In: Saerens, D., Muyldermans, S. (eds) Single Domain Antibodies. Methods in Molecular Biology, vol 911. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-968-6_5
Download citation
DOI: https://doi.org/10.1007/978-1-61779-968-6_5
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-967-9
Online ISBN: 978-1-61779-968-6
eBook Packages: Springer Protocols