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

TitleGeneration of single domain antibody fragments derived from camelids and generation of manifold constructs.
Publication TypeJournal Article
Year of Publication2012
AuthorsVincke, C., C. GutiƩrrez, U. Wernery, N. Devoogdt, G. Hassanzadeh-Ghassabeh, and S. Muyldermans
JournalMethods Mol Biol
Volume907
Pagination145-76
Date Published2012
ISSN1940-6029
KeywordsAmino Acid Sequence, Animals, Bacteriophages, Base Sequence, Camelids, New World, Camels, DNA, Complementary, Electroporation, Epitopes, Escherichia coli, Genetic Vectors, Immunization, Immunoglobulin Heavy Chains, Lymphocytes, Molecular Biology, Molecular Sequence Data, Polymerase Chain Reaction, Restriction Mapping, RNA, Single-Domain Antibodies
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.

DOI10.1007/978-1-61779-974-7_8
Alternate JournalMethods Mol. Biol.
PubMed ID22907350