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. 2012:907:145-76.
doi: 10.1007/978-1-61779-974-7_8.

Generation of single domain antibody fragments derived from camelids and generation of manifold constructs

Cécile Vincke  1 , Carlos Gutiérrez, Ulrich Wernery, Nick Devoogdt, Gholamreza Hassanzadeh-Ghassabeh, Serge Muyldermans
Affiliations

Generation of single domain antibody fragments derived from camelids and generation of manifold constructs

Cécile Vincke et al. Methods Mol Biol. 2012.

Abstract

Immunizing a camelid (camels and llamas) with soluble, properly folded proteins raises an affinity-matured immune response in the unique camelid heavy-chain only antibodies (HCAbs). The peripheral blood lymphocytes of the immunized animal are used to clone the antigen-binding antibody fragment from the HCAbs in a phage display vector. A representative aliquot of the library of these antigen-binding fragments is used to retrieve single domain antigen-specific binders by successive rounds of panning. These single domain antibody fragments are cloned in tandem to generate manifold constructs (bivalent, biparatopic or bispecific constructs) to increase their functional affinity, to increase specificity, or to connect two independent antigen molecules.

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