title: Guided selection of antibody fragments specific for human interferon gamma receptor 1 from a human VH- and VL-gene repertoire.
abstract: OBJECTIVE
  The guided selection strategy for isolation of human antibody (Ab) fragments specific for human interferon gamma receptor 1 (IFNGR-1) from a cloned Ab VH and VL repertoire has been investigated. In order to identify recombinant Abs binding to soluble antigen, a novel method termed affinity sedimentation was introduced here.
  
  
  CONCLUSIONS
  The VH region of murine monoclonal Ab (IR gamma-1) against human IFNGR-1 was combined with human VL repertoire and used for selection of human VL regions. One of these human VL regions (kappa 2) possesses high homology to the murine template VL region, also in CDR3 (77%). A chimeric Fab consisting of kappa 2 and the murine IR gamma-1 VH region was highly IFNGR-1 specific and exerted the same epitope specificity and a comparable binding affinity as the parental murine Fab. In a further step, the selected human VL region kappa 2 was combined with a human VH repertoire and led by guided selection to the generation of a completely human Fab (1b5) specific for human IFNGR-1. The overall VH region homology of 1b5 compared to the parental antibody IR gamma-1 was 81%, with a rather low homology in CDR3. Binding competition studies revealed that the epitope recognized by 1b5 differs from the parental Ab IR gamma-1.
authors.affiliation: Institute of Cell Biology and Immunology, University of Stuttgart, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Watzka
authors.forename: K
authors.initials: K
authors.lastname: Pfizenmaier
authors.forename: D
authors.initials: D
authors.lastname: Moosmayer
mesh_terms: D000595:Amino Acid Sequence; D000906:Antibodies / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D000915:Antibody Affinity / Q000235:genetics; D000918:Antibody Specificity / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D005803:Genes, Immunoglobulin; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics* / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000235:genetics* / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D017471:Receptors, Interferon / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D000096743:Interferon gamma Receptor
chemical_list: D000906:Antibodies; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D017471:Receptors, Interferon; D011994:Recombinant Proteins; D000096743:Interferon gamma Receptor