title: Generation of anti-infectome/anti-proteome nanobodies.
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.
authors.affiliation: Department of Molecular and Cellular Interactions, VIB, Brussels, Belgium. ghassanz@vub.ac.be
authors.forename: Gholamreza
authors.initials: G
authors.lastname: Hassanzadeh-Ghassabeh
authors.forename: Dirk
authors.initials: D
authors.lastname: Saerens
authors.forename: Serge
authors.initials: S
authors.lastname: Muyldermans
mesh_terms: D000818:Animals; D000918:Antibody Specificity; D002846:Chromatography, Affinity; D004262:DNA Restriction Enzymes / Q000378:metabolism; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics; D007128:Immunoglobulin Fragments / Q000737:chemistry* / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D007963:Leukocytes, Mononuclear / Q000276:immunology; D049329:Nanostructures; D019151:Peptide Library; D019897:Periplasm / Q000378:metabolism; D016133:Polymerase Chain Reaction; D020543:Proteome / Q000276:immunology*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D014345:Trypanosoma / Q000276:immunology*
chemical_list: D018076:DNA, Complementary; D007128:Immunoglobulin Fragments; D019151:Peptide Library; D020543:Proteome; D011994:Recombinant Proteins; D012313:RNA; D004262:DNA Restriction Enzymes