Design, construction, and characterization of a large synthetic human antibody phage display library

Proteomics. 2005 Jun;5(9):2340-50. doi: 10.1002/pmic.200401273.


Advances in proteomic research allow the identification of several hundred protein components in complex biological specimens. Structural information is typically lost during proteomic investigations. For this reason, the rapid isolation of monoclonal antibodies specific to proteins of interest would allow the study of structurally intact biological specimens, thus providing complementary proteomic information. Here, we describe the design, construction, characterization, and use of a large synthetic human antibody phage display library (ETH-2-Gold) containing three billion individual antibody clones. A large repertoire of antibodies with similar biochemical properties was produced by appending short variable complementarity-determining region 3 (CDR3) onto three antibody germline segments (DP47, DPK22, and DPL16), which are frequently found in human antibodies. The ETH-2-Gold library exhibits efficient display of antibody fragments on filamentous phage, as assessed by immunoblot. Furthermore, the library is highly functional, since >90% of clones express soluble antibodies in bacteria and since good quality monoclonal antibodies have been isolated against 16 different antigens. The usefulness of the library as a tool for generating monoclonal antibodies for biomedical applications was tested using the C-domain of tenascin-C (a marker of angiogenesis) as antigen and showing that specific antibodies to this target were able to stain vascular structures in tumor sections.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal* / biosynthesis
  • Antibodies, Monoclonal* / genetics
  • Antibody Specificity
  • Bacteria / metabolism
  • Capsid Proteins
  • Cloning, Molecular
  • DNA-Binding Proteins / genetics
  • Enzyme-Linked Immunosorbent Assay
  • Glioblastoma / blood supply
  • Humans
  • Immunoglobulin Variable Region / genetics
  • Inovirus / metabolism
  • Neovascularization, Pathologic / metabolism
  • Peptide Library*
  • Protein Structure, Tertiary
  • Tenascin / analysis
  • Tenascin / immunology
  • Viral Fusion Proteins / genetics


  • Antibodies, Monoclonal
  • Capsid Proteins
  • DNA-Binding Proteins
  • Immunoglobulin Variable Region
  • Peptide Library
  • Tenascin
  • Viral Fusion Proteins