Affinity maturation and functional dissection of a humanised anti-RAGE monoclonal antibody by ribosome display

Methods Mol Biol. 2012:805:403-22. doi: 10.1007/978-1-61779-379-0_23.

Abstract

The pursuit of more potent, safe, and cost-effective drugs has placed a greater emphasis on antibody optimisation within the drug discovery process. Technologies to rapidly improve antibody drug performance, such as phage display, ribosome display, and yeast display, are playing a key role in this effort. Among these ribosome display is a particularly powerful technology and has recently been applied to the affinity optimisation of a humanised anti-receptor for advanced glycation end products (anti-RAGE) antibody (Finlay et al., J Mol Biol 388:541-558, 2009). By using a combination of error-prone PCR with ribosome display each amino acid position within this humanised antibody was scanned for both its functional importance and its capacity to increase affinity resulting in both affinity-matured antibody variants and a functional map of the antibody paratope.

MeSH terms

  • Antibodies, Monoclonal / genetics*
  • Antibodies, Monoclonal / immunology*
  • Antibody Affinity*
  • Directed Molecular Evolution*
  • Glycation End Products, Advanced / immunology*
  • Humans
  • Polymerase Chain Reaction
  • Ribosomes / genetics*

Substances

  • Antibodies, Monoclonal
  • Glycation End Products, Advanced