Generation of high-affinity human antibodies by combining donor-derived and synthetic complementarity-determining-region diversity

Nat Biotechnol. 2005 Mar;23(3):344-8. doi: 10.1038/nbt1067. Epub 2005 Feb 20.

Abstract

Combinatorial libraries of rearranged hypervariable V(H) and V(L) sequences from nonimmunized human donors contain antigen specificities, including anti-self reactivities, created by random pairing of V(H)s and V(L)s. Somatic hypermutation of immunoglobulin genes, however, is critical in the generation of high-affinity antibodies in vivo and occurs only after immunization. Thus, in combinatorial phage display libraries from nonimmunized donors, high-affinity antibodies are rarely found. Lengthy in vitro affinity maturation is often needed to improve antibodies from such libraries. We report the construction of human Fab libraries having a unique combination of immunoglobulin sequences captured from human donors and synthetic diversity in key antigen contact sites in heavy-chain complementarity-determining regions 1 and 2. The success of this strategy is demonstrated by identifying many monovalent Fabs against multiple therapeutic targets that show higher affinities than approved therapeutic antibodies. This very often circumvents the need for affinity maturation, accelerating discovery of antibody drug candidates.

Publication types

  • Letter

MeSH terms

  • Antibody Affinity*
  • Antibody Formation*
  • Complementarity Determining Regions / genetics*
  • Genetic Variation / genetics
  • Humans
  • Immunoglobulin Fab Fragments / biosynthesis*
  • Immunoglobulin Fab Fragments / genetics
  • Immunoglobulin Fab Fragments / immunology*
  • Peptide Library*
  • Protein Binding
  • Protein Engineering / methods*
  • Recombination, Genetic / genetics
  • Tissue Donors

Substances

  • Complementarity Determining Regions
  • Immunoglobulin Fab Fragments
  • Peptide Library