Generation of anti-infectome/anti-proteome nanobodies.

TitleGeneration of anti-infectome/anti-proteome nanobodies.
Publication TypeJournal Article
Year of Publication2011
AuthorsHassanzadeh-Ghassabeh, G., D. Saerens, and S. Muyldermans
JournalMethods Mol Biol
Volume790
Pagination239-59
Date Published2011
ISSN1940-6029
KeywordsAnimals, Antibody Specificity, Chromatography, Affinity, DNA Restriction Enzymes, DNA, Complementary, Escherichia coli, Immunoglobulin Fragments, Leukocytes, Mononuclear, Nanostructures, Peptide Library, Periplasm, Polymerase Chain Reaction, Proteome, Recombinant Proteins, RNA, Trypanosoma
Abstract

The immunization of an animal with a whole proteome or the infection of an animal and the screening of the resulting antibody repertoire on either the same or different proteome(s) or the infecting agent(s), omits the laborious steps of recombinant protein expression and purification to obtain multiple antigen binders. This procedure allows the identification of antibodies that are specific to unique or common signatures of different proteomes without prior knowledge of these signatures.Nanobodies are the smallest (15 kDa, 2.2 nm diameter, 4 nm height) in vivo affinity-matured functional antigen-binding entities that are derived from camelid heavy-chain antibodies. Due to their small size, recognition of unique epitopes, high affinity, and easy tailoring, nanobodies are attractive affinity reagents for various applications, including diagnosis and therapy.We detail a protocol to generate, isolate, express, and purify anti-infectome/anti-proteome nanobodies.

DOI10.1007/978-1-61779-319-6_19
Alternate JournalMethods Mol. Biol.
PubMed ID21948420