Selection of human antibody fragments by phage display.
Here, we describe a protocol for the selection of human antibody fragments using repertoires displayed on filamentous bacteriophage. Antigen-specific clones are enriched by binding to immobilized antigen, followed by elution and repropagation of phage. After multiple rounds of binding selection, specific clones are identified by ELISA. This article provides an overview of phage display and antibody technology, as well as detailed protocols for the immobilization of antigen, the selection of repertoires on purified or complex antigens and the identification of binders.
Lee
Carol M Y
CM
Garvan Institute of Medical Research, 384 Victoria Road, Darlinghurst, Sydney New South Wales 2010, Australia.
Iorno
Niccolo
N
Sierro
Frederic
F
Christ
Daniel
D
D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000378:metabolism; D000937:Antigen-Antibody Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007128:Immunoglobulin Fragments / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D019151:Peptide Library; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D019151:Peptide Library; D011993:Recombinant Fusion Proteins