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Research article

References

  1. Bidlingmaier S, He J, Wang Y, An F, Feng J, Barbone D, et al. 2009. Identification of MCAM/CD146 as the target antigen of a human monoclonal antibody that recognizes both epithelioid and sarcomatoid types of mesothelioma. Cancer Res. 69: 1570-1577.
    Pubmed PMC CrossRef
  2. Bidlingmaier S, Liu B. 2006. Construction and application of a yeast surface-displayed human cDNA library to identify post-translational modification-dependent protein-protein interactions. Mol. Cell. Proteomics 5: 533-540.
    Pubmed CrossRef
  3. Bidlingmaier S, Liu B. 2011. Construction of yeast surfacedisplayed cDNA libraries. Methods Mol. Biol. 729: 199-210.
    Pubmed PMC CrossRef
  4. Chao G, Lau WL, Hackel BJ, Sazinsky SL, Lippow SM, Wittrup KD. 2006. Isolating and engineering human antibodies using yeast surface display. Nat. Protoc. 1: 755-768.
    Pubmed CrossRef
  5. Daugherty PS, Iverson BL, Georgiou G. 2000. Flow cytometric screening of cell-based libraries. J. Immunol. Methods 243:211-227.
    CrossRef
  6. Davis C, Barvish Z, Gitelman I. 2007. A method for the construction of equalized directional cDNA libraries from hydrolyzed total RNA. BMC Genomics 8: 363.
    Pubmed PMC CrossRef
  7. Foung S, Perkins S, Kafadar K, Gessner P, Zimmermann U. 1990. Development of microfusion techniques to generate human hybridomas. J. Immunol. Methods 134: 35-42.
    CrossRef
  8. Foung SK, Perkins S. 1989. Electric field-induced cell fusion and human monoclonal antibodies. J. Immunol. Methods 116:117-122.
    CrossRef
  9. Galluzzi L, Vacchelli E, Fridman WH, Galon J, SautsFridman C, Tartour E, et al. 2012. Trial watch: monoclonal antibodies in cancer therapy. Oncoimmunology 1: 28.
    Pubmed PMC CrossRef
  10. Howland SW, Poh C-M, Rnia L. 2011. Directional, seamless, and restriction enzyme-free construction of random-primed complementary DNA libraries using phosphorothioatemodified primers. Anal. Biochem. 416: 141-143.
    Pubmed CrossRef
  11. Jack HM, Beck-Engeser G, Lee G, Wofsy D, Wabl M. 1992. Tumorigenesis mediated by an antigen receptor. Proc. Natl. Acad. Sci. USA 89: 8482-8486.
    Pubmed PMC CrossRef
  12. Jung V, Fischer E, Imig J, Kleber S, Nuber N, Reinshagen F, et al. 2010. Yeast-based identification of prostate tumor antigens provides an effective vaccine platform. Anticancer Res. 30: 895-902.
    Pubmed
  13. Kieke MC, Cho BK, Boder ET, Kranz DM, Wittrup KD. 1997. Isolation of anti-T cell receptor scFv mutants by yeast surface display. Protein Eng. 10: 1303-1310.
    Pubmed CrossRef
  14. Kim M, Choi HY, Choi YS, Kim J, Kim Y. 2007. Optimized serological isolation of lung-cancer-associated antigens from a yeast surface-expressed cDNA library. J. Microbiol. Biotechnol. 17: 993.
    Pubmed
  15. Kondo A, Ueda M. 2004. Yeast cell-surface display −applications of molecular display. Appl. Microbiol. Biotechnol. 64: 28-40.
    Pubmed CrossRef
  16. Pepper LR, Cho YK, Boder ET, Shusta EV. 2008. A decade of yeast surface display technology: where are we now? Comb. Chem. High Throughput Screen. 11: 127.
    Pubmed PMC CrossRef
  17. Van Antwerp JJ, Wittrup KD. 2000. Fine affinity discrimination by yeast surface display and flow cytometry. Biotechnol. Prog. 16: 31-37.
    Pubmed CrossRef
  18. Wadle A, Mischo A, Imig J, Wllner B, Hensel D, Wtzig K, et al. 2005. Serological identification of breast cancer-related antigens from a Saccharomyces cerevisiae surface display library. Int. J. Cancer 117: 104-113.
    Pubmed CrossRef
  19. Yeung YA, Wittrup KD. 2002. Quantitative screening of yeast surface-displayed polypeptide libraries by magnetic bead capture. Biotechnol. Prog. 18: 212-220.
    Pubmed CrossRef
  20. Yu X, McGraw PA, House FS, Crowe JE Jr. 2008. An optimized electrofusion-based protocol for generating virusspecific human monoclonal antibodies. J. Immunol. Methods 336: 142-151.
    Pubmed PMC CrossRef

Article

Research article

J. Microbiol. Biotechnol. 2015; 25(4): 547-553

Published online April 28, 2015 https://doi.org/10.4014/jmb.1409.09079

Copyright © The Korean Society for Microbiology and Biotechnology.

Optimization of Yeast Surface-Displayed cDNA Library Screening for Low Abundance Targets

Juhyung Kim 1, Hyung Kyu Kim 1, Hye Jeong Jang 1, Eunkyung Kim 1 and Moon Kyu Kim 1*

Department of Immunology, School of Medicine, Kyungpook National University, Daegu 700-422, Republic of Korea

Received: September 25, 2014; Accepted: November 10, 2014

Abstract

The yeast surface-displayed cDNA library has been used to identify unknown antigens.
However, when unknown target antigens show moderate-to-low abundance, some
modifications are needed in the screening process. In this study, a directional random-primed
cDNA library was used to increase the number of candidates for the unknown antigen. To
avoid the loss of target yeast clones that express proteins at a low frequency in the cDNA
library, a comprehensive monitoring system based on magnetic-activated cell sorting,
fluorescence-activated cell sorting, and immunofluorescence was established, and a small
number of target yeast cells was successfully enriched. These results showed that our
optimized method has potential application for identifying rare unknown antigens of the
human monoclonal antibody.

Keywords: cDNA library, yeast surface display, tumor-associated antigen, hepatocellular carcinoma

References

  1. Bidlingmaier S, He J, Wang Y, An F, Feng J, Barbone D, et al. 2009. Identification of MCAM/CD146 as the target antigen of a human monoclonal antibody that recognizes both epithelioid and sarcomatoid types of mesothelioma. Cancer Res. 69: 1570-1577.
    Pubmed KoreaMed CrossRef
  2. Bidlingmaier S, Liu B. 2006. Construction and application of a yeast surface-displayed human cDNA library to identify post-translational modification-dependent protein-protein interactions. Mol. Cell. Proteomics 5: 533-540.
    Pubmed CrossRef
  3. Bidlingmaier S, Liu B. 2011. Construction of yeast surfacedisplayed cDNA libraries. Methods Mol. Biol. 729: 199-210.
    Pubmed KoreaMed CrossRef
  4. Chao G, Lau WL, Hackel BJ, Sazinsky SL, Lippow SM, Wittrup KD. 2006. Isolating and engineering human antibodies using yeast surface display. Nat. Protoc. 1: 755-768.
    Pubmed CrossRef
  5. Daugherty PS, Iverson BL, Georgiou G. 2000. Flow cytometric screening of cell-based libraries. J. Immunol. Methods 243:211-227.
    CrossRef
  6. Davis C, Barvish Z, Gitelman I. 2007. A method for the construction of equalized directional cDNA libraries from hydrolyzed total RNA. BMC Genomics 8: 363.
    Pubmed KoreaMed CrossRef
  7. Foung S, Perkins S, Kafadar K, Gessner P, Zimmermann U. 1990. Development of microfusion techniques to generate human hybridomas. J. Immunol. Methods 134: 35-42.
    CrossRef
  8. Foung SK, Perkins S. 1989. Electric field-induced cell fusion and human monoclonal antibodies. J. Immunol. Methods 116:117-122.
    CrossRef
  9. Galluzzi L, Vacchelli E, Fridman WH, Galon J, SautsFridman C, Tartour E, et al. 2012. Trial watch: monoclonal antibodies in cancer therapy. Oncoimmunology 1: 28.
    Pubmed KoreaMed CrossRef
  10. Howland SW, Poh C-M, Rnia L. 2011. Directional, seamless, and restriction enzyme-free construction of random-primed complementary DNA libraries using phosphorothioatemodified primers. Anal. Biochem. 416: 141-143.
    Pubmed CrossRef
  11. Jack HM, Beck-Engeser G, Lee G, Wofsy D, Wabl M. 1992. Tumorigenesis mediated by an antigen receptor. Proc. Natl. Acad. Sci. USA 89: 8482-8486.
    Pubmed KoreaMed CrossRef
  12. Jung V, Fischer E, Imig J, Kleber S, Nuber N, Reinshagen F, et al. 2010. Yeast-based identification of prostate tumor antigens provides an effective vaccine platform. Anticancer Res. 30: 895-902.
    Pubmed
  13. Kieke MC, Cho BK, Boder ET, Kranz DM, Wittrup KD. 1997. Isolation of anti-T cell receptor scFv mutants by yeast surface display. Protein Eng. 10: 1303-1310.
    Pubmed CrossRef
  14. Kim M, Choi HY, Choi YS, Kim J, Kim Y. 2007. Optimized serological isolation of lung-cancer-associated antigens from a yeast surface-expressed cDNA library. J. Microbiol. Biotechnol. 17: 993.
    Pubmed
  15. Kondo A, Ueda M. 2004. Yeast cell-surface display −applications of molecular display. Appl. Microbiol. Biotechnol. 64: 28-40.
    Pubmed CrossRef
  16. Pepper LR, Cho YK, Boder ET, Shusta EV. 2008. A decade of yeast surface display technology: where are we now? Comb. Chem. High Throughput Screen. 11: 127.
    Pubmed KoreaMed CrossRef
  17. Van Antwerp JJ, Wittrup KD. 2000. Fine affinity discrimination by yeast surface display and flow cytometry. Biotechnol. Prog. 16: 31-37.
    Pubmed CrossRef
  18. Wadle A, Mischo A, Imig J, Wllner B, Hensel D, Wtzig K, et al. 2005. Serological identification of breast cancer-related antigens from a Saccharomyces cerevisiae surface display library. Int. J. Cancer 117: 104-113.
    Pubmed CrossRef
  19. Yeung YA, Wittrup KD. 2002. Quantitative screening of yeast surface-displayed polypeptide libraries by magnetic bead capture. Biotechnol. Prog. 18: 212-220.
    Pubmed CrossRef
  20. Yu X, McGraw PA, House FS, Crowe JE Jr. 2008. An optimized electrofusion-based protocol for generating virusspecific human monoclonal antibodies. J. Immunol. Methods 336: 142-151.
    Pubmed KoreaMed CrossRef