Generation of human Fab libraries for phage display.
This protocol describes the generation of human antibody libraries in Fab format from 2.5 × 10(7) human peripheral blood or bone marrow mononuclear cells for their subsequent selection by phage display. Although it can be applied to the mining of both human naïve and immune antibody repertoires, the procedure is primarily intended for the generation of fully human monoclonal antibodies from patients with endogenous antibody responses of interest and limited availability of clinical specimens.
Experimental Transplantation and Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. raderc@mail.nih.gov
Christoph
C
Rader
D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D000915:Antibody Affinity / Q000235:genetics / Q000276:immunology; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D019151:Peptide Library; D016133:Polymerase Chain Reaction
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library