Directed evolution for improved secretion of cancer–testis antigen NY-ESO-1 from yeast Academic Article uri icon


  • NY-ESO-1 is a highly immunogenic tumor antigen and a promising vaccine candidate in cancer immunotherapy. Access to purified protein both for vaccine formulations and for monitoring antigen-specific immune responses is vital to vaccine development. Currently available recombinant Escherichia coli-derived NY-ESO-1 is isolated from inclusion bodies as a complex protein mixture and efforts to improve the purity of this antigen are required, especially for later-stage clinical trials. Using yeast cell surface display and fluorescence activated cell sorting techniques, we have engineered an NY-ESO-1 variant (NY-ESO-L5; C(75)A C(76)A C(78)A L(153)H) with a 100x improved display level on yeast compared to the wild-type protein. This mutant can be effectively produced as an Aga2p-fusion and purified in soluble form directly from the yeast cell wall. In the process, we have identified the epitope recognized by anti-NY-ESO-1 mAb E978 (79-87, GARGPESRL). The availability of an alternative expression host for this important antigen will help avoid artifactual false positive tests of patient immune response due to reaction against expression-host-specific contaminants.


  • Piatesi, Andrea
  • Howland, Shanshan W
  • Rakestraw, James A
  • Renner, Christoph
  • Robson, Neil
  • Cebon, Jonathan
  • Maraskovsky, Eugene
  • Ritter, Gerd
  • Old, Lloyd
  • Wittrup, K Dane

publication date

  • August 2006