Generation of single domain antibody fragments derived from camelids and generation of manifold constructs.
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.
Department of Structural Biology, VIB, Vrije Universiteit Brussel, Brussel, Belgium.
Cécile
C
Vincke
Carlos
C
Gutiérrez
Ulrich
U
Wernery
Nick
N
Devoogdt
Gholamreza
G
Hassanzadeh-Ghassabeh
Serge
S
Muyldermans
D000595:Amino Acid Sequence; D000818:Animals; D001435:Bacteriophages / Q000378:metabolism; D001483:Base Sequence; D002161:Camelids, New World / Q000276:immunology*; D002162:Camelus / Q000276:immunology*; D018076:DNA, Complementary / Q000096:biosynthesis; D018274:Electroporation; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli; D005822:Genetic Vectors / Q000235:genetics; D007114:Immunization; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D008214:Lymphocytes / Q000378:metabolism; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012313:RNA / Q000378:metabolism; D015183:Restriction Mapping; D061905:Single-Domain Antibodies / Q000235:genetics*
D018076:DNA, Complementary; D000939:Epitopes; D007143:Immunoglobulin Heavy Chains; D061905:Single-Domain Antibodies; D012313:RNA