title: Construction of a human antibody domain (VH) library.
abstract: Highly diverse antibody (Fab or scFv) libraries have become vital sources to select antibodies with high affinity and novel properties. Combinatorial strategies provide efficient ways of creating antibody libraries containing a large number of individual clones. These strategies include the reassembly of naturally occurring genes encoding the heavy and light chains from either immune or nonimmune B-cell sources, or introduction of synthetic diversity to either the framework regions (FRs) or the complementarity-determining regions (CDRs) of the variable domains of antibodies. In the late 1980s, the smallest known antigen-binding fragment was identified when a murine VH repertoire was screened for binding to lysozyme. This fragment (approximately 15 kDa), called a "domain antibody", or "dAb", is approximately four times smaller than a Fab and half the size of a scFv. Here, we describe the construction of a phage-displayed VH library and an approach to introduce genetic diversity in this library, where both diverse human CDRs and synthetic CDRs are combined into a single-domain (VH) framework.
authors.affiliation: NCI-Frederick, National Institutes of Health, Frederick, MD, USA.
authors.forename: Weizao
authors.initials: W
authors.lastname: Chen
authors.forename: Zhongyu
authors.initials: Z
authors.lastname: Zhu
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Xiao
authors.forename: Dimiter S
authors.initials: DS
authors.lastname: Dimitrov
mesh_terms: D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D002469:Cell Separation; D022801:Complementarity Determining Regions / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D018274:Electroporation; D005822:Genetic Vectors; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D017434:Protein Structure, Tertiary; D012313:RNA / Q000302:isolation & purification; D014158:Transcription, Genetic
chemical_list: D000906:Antibodies; D022801:Complementarity Determining Regions; D018076:DNA, Complementary; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D012313:RNA