Optimized Serological Isolation of Lung-Cancer-associated Antigens from a Yeast Surface-expressed cDNA Library

  • Kim, Min-Soo (Department of Molecular Science and Technology, Ajou University) ;
  • Choi, Hye-Young (Cancer Research Center, Center for Clinical Research, Samsung Biomedical Research Institute) ;
  • Choi, Yong-Soo (Department of Thoracic and Cardiovascular Surgery, Samsung Medical Center, Sungkyunkwan University, School of Medicine) ;
  • Kim, Jhin-Gook (Department of Thoracic and Cardiovascular Surgery, Samsung Medical Center, Sungkyunkwan University, School of Medicine) ;
  • Kim, Yong-Sung (Department of Molecular Science and Technology, Ajou University)
  • Published : 2007.06.30

Abstract

The technique of serological analysis of antigens by recombinant cDNA expression library (SEREX) uses autologous patient sera as a screening probe to isolate tumor-associated antigens for various tumor types. Isolation of tumor-associated antigens that are specifically reactive with patient sera, but not with normal sera, is important to avoid false-positive and autoimmunogenic antigens for the cancer immunotherapy. Here, we describe a selection methodology to isolate patient sera-specific antigens from a yeast surface-expressed cDNA library constructed from 15 patient lung tissues with non-small cell lung cancer (NSCLC). Several rounds of positive selection using patient sera alone as a screening probe isolated clones exhibiting comparable reactivity with both patient and normal sera. However, the combination of negative selection with allogeneic normal sera to remove antigens reactive with normal sera and subsequent positive selection with patient sera efficiently enriched patient sera-specific antigens. Using the selection methodology described here, we isolated 3 known and 5 unknown proteins, which have not been isolated previously, but and potentially associated with NSCLC.

Keywords

References

  1. Bielas, J. H., K. R. Loeb, B. P. Rubin, L. D. True, and L. A. Loeb. 2006. From the cover: Human cancers express a mutator phenotype. Proc. Natl. Acad. Sci. USA 103: 18238-18242
  2. Cha, K. H., M. D. Kim, T. H. Lee, H. K. Lim, K. H. Jung, and J. H. Seo. 2006. Coexpression of protein disulfide isomerase (PDI) enhances production of kringle fragment of human apolipoprotein(a) in recombinant Saccharomyces cerevisiae. J. Microbiol. Biotechnol. 16: 308-311
  3. Colby, D. W., B. A. Kellogg, C. P. Graff, Y. A. Yeung, J. S. Swers, and K. D. Wittrup. 2004. Engineering antibody affinity by yeast surface display. Methods Enzymol. 388: 348-358 https://doi.org/10.1016/S0076-6879(04)88027-3
  4. Goldsby, R. A., T. J. Kindt, B. A. Osborne, and J. Kuby. 2003. Immunology, 5th Ed. Freeman, New York
  5. Guinn, B. A., J. F. Collin, G. Li, R. C. Rees, and G. J. Mufti. 2002. Optimised SEREX technique for the identification of leukaemiaassociated antigens. J. Immunol. Methods 264: 207-214 https://doi.org/10.1016/S0022-1759(02)00095-9
  6. Gure, A. O., E. Stockert, M. J. Scanlan, R. S. Keresztes, D. Jager, N. K. Altorki, L. J. Old, and Y. T. Chen. 2000. Serological identification of embryonic neural proteins as highly immunogenic tumor antigens in small cell lung cancer. Proc. Natl. Acad. Sci. USA 97: 4198-4203
  7. Gure, A. O., N. K. Altorki, E. Stockert, M. J. Scanlan, L. J. Old, and Y. T. Chen. 1998. Human lung cancer antigens recognized by autologous antibodies: Definition of a novel cDNA derived from the tumor suppressor gene locus on chromosome 3p21.3. Cancer Res. 58: 1034-1041
  8. Honda, K., T. Yamada, R. Endo, Y. Ino, M. Gotoh, H. Tsuda, Y. Yamada, H. Chiba, and S. Hirohashi. 1998. Actinin-4, a novel actin-bundling protein associated with cell motility and cancer invasion. J. Cell Biol. 140: 1383-1393 https://doi.org/10.1083/jcb.140.6.1383
  9. Jager, E., S. Gnjatic, Y. Nagata, E. Stockert, D. Jager, J. Karbach, A. Neumann, J. Rieckenberg, Y. T. Chen, G. Ritter, E. Hoffman, M. Arand, L. J. Old, and A. Knuth. 2000. Induction of primary NY-ESO-1 immunity: CD8+ T lymphocyte and antibody responses in peptide-vaccinated patients with NYESO- 1+ cancers. Proc. Natl. Acad. Sci. USA 97: 12198-12203
  10. Kang, J. S., M. Gao, J. L. Feinleib, P. D. Cotter, S. N. Guadagno, and R. S. Krauss. 1997. CDO: An oncogene-, serum-, and anchorage-regulated member of the Ig/fibronectin type III repeat family. J. Cell Biol. 138: 203-213 https://doi.org/10.1083/jcb.138.1.203
  11. Kim, H. C., H. J. Kim, W. B. Choi, and S. W. Nam. 2006. Inulooligosaccharide production from inulin by Saccharomyces cerevisiae strain displaying cell-surface endoinulinase. J. Microbiol. Biotechnol. 16: 360-367
  12. Kim, Y. S., R. Bhandari, J. R. Cochran, J. Kuriyan, and K. D. Wittrup. 2006. Directed evolution of the epidermal growth factor receptor extracellular domain for expression in yeast. Proteins 62: 1026-1035 https://doi.org/10.1002/prot.20618
  13. Lee, H. W., S. H. Lee, K. J. Park, J. S. Kim, M. H. Kwon, and Y. S. Kim. 2006. Construction and characterization of a pseudo-immune human antibody library using yeast surface display. Biochem. Biophys. Res. Commun. 346: 896-903 https://doi.org/10.1016/j.bbrc.2006.05.202
  14. Li, G., A. Miles, A. Line, and R. C. Rees. 2004. Identification of tumour antigens by serological analysis of cDNA expression cloning. Cancer Immunol. Immunother. 53: 139-143 https://doi.org/10.1007/s00262-003-0471-y
  15. Mischo, A., A. Wadle, K. Watzig, D. Jager, E. Stockert, D. Santiago, G. Ritter, E. Regitz, E. Jager, A. Knuth, L. Old, M. Pfreundschuh, and C. Renner. 2003. Recombinant antigen expression on yeast surface (RAYS) for the detection of serological immune responses in cancer patients. Cancer Immun. 3: 5
  16. Preuss, K. D., C. Zwick, C. Bormann, F. Neumann, and M. Pfreundschuh. 2002. Analysis of the B-cell repertoire against antigens expressed by human neoplasms. Immunol. Rev. 188: 43-50 https://doi.org/10.1034/j.1600-065X.2002.18805.x
  17. Sahin, U., O. Tureci, H. Schmitt, B. Cochlovius, T. Johannes, R. Schmits, F. Stenner, G. Luo, I. Schobert, and M. Pfreundschuh. 1995. Human neoplasms elicit multiple specific immune responses in the autologous host. Proc. Natl. Acad. Sci. USA 92: 11810-11813
  18. Siegel, R. W., J. R. Coleman, K. D. Miller, and M. J. Feldhaus. 2004. High efficiency recovery and epitopespecific sorting of an scFv yeast display library. J. Immunol. Methods 286: 141-153 https://doi.org/10.1016/j.jim.2004.01.005
  19. Tureci, O., U. Sahin, C. Zwick, M. Koslowski, G. Seitz, and M. Pfreundschuh. 1998. Identification of a meiosis-specific protein as a member of the class of cancer/testis antigens. Proc. Natl. Acad. Sci. USA 95: 5211-5216
  20. Tureci, O., U. Sahin, and M. Pfreundschuh. 1997. Serological analysis of human tumor antigens: Molecular definition and implications. Mol. Med. Today 3: 342-349 https://doi.org/10.1016/S1357-4310(97)01081-2
  21. Vuillier, F., G. Dumas, C. Magnac, M. C. Prevost, A. I. Lalanne, P. Oppezzo, E. Melanitou, G. Dighiero, and B. Payelle- Brogard. 2005. Lower levels of surface B-cell-receptor expression in chronic lymphocytic leukemia are associated with glycosylation and folding defects of the mu and CD79a chains. Blood 105: 2933-2940 https://doi.org/10.1182/blood-2004-09-3643
  22. Wadle, A., A. Mischo, J. Imig, B. Wullner, D. Hensel, K. Watzig, F. Neumann, B. Kubuschok, W. Schmidt, L. J. Old, M. Pfreundschuh, and C. Renner. 2005. Serological identification of breast cancer-related antigens from a Saccharomyces cerevisiae surface display library. Int. J. Cancer 117: 104-113 https://doi.org/10.1002/ijc.21147
  23. Wadle, A., B. Kubuschok, J. Imig, B. Wuellner, C. Wittig, C. Zwick, A. Mischo, K. Waetzig, B. F. Romeike, W. Lindemann, M. Schilling, M. Pfreundschuh, and C. Renner. 2006. Serological immune response to cancer testis antigens in patients with pancreatic cancer. Int. J. Cancer 119: 117-125 https://doi.org/10.1002/ijc.21744
  24. Yajnik, V., C. Paulding, R. Sordella, A. I. McClatchey, M. Saito, D. C. Wahrer, P. Reynolds, D. W. Bell, R. Lake, S. van den Heuvel, J. Settleman, and D. A. Haber. 2003. DOCK4, a GTPase activator, is disrupted during tumorigenesis. Cell 112: 673-684 https://doi.org/10.1016/S0092-8674(03)00155-7
  25. Zheng, B., Y. C. Ma, R. S. Ostrom, C. Lavoie, G. N. Gill, P. A. Insel, X. Y. Huang, and M. G. Farquhar. 2001. RGSPX1, a GAP for GalphaS and sorting nexin in vesicular trafficking. Science 294: 1939-1942 https://doi.org/10.1126/science.1064757