Skip to main content
SpringerLink
Log in

Synthesis and Cell-Based Screening of One-Bead-One-Compound Peptide Libraries

  • Protocol
  • First Online:
Peptide Libraries

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1248))

Abstract

Combinatorial one-bead-one-compound (OBOC) peptide library screening has proven to be a powerful tool for identification of small molecules, peptides, or peptidomimetics against a variety of specific targets such as cell surface receptors, protein kinases, proteases, and phosphatases. With each bead displaying many copies of a single chemical entity, millions of compounds can be rapidly synthesized and screened with whole-cell binding on-bead functional assays. Here we describe the methodology for the synthesis, screening, and sequence deconvolution of an OBOC peptide library analyzed for affinity to a cancer cell line.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Geysen HM, Meloen RB, Barteling SJ (1984) Use of peptide synthesis to probe viral antigens for epitopes to a resolution of a single amino acid. Proc Natl Acad Sci 81:3998–4002

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  2. Lam KS, Salmon SE, Hersh EM, Hruby VJ, Kazmierski WM, Knapp RJ (1991) A new type of synthetic peptide library for identifying ligand-binding activity. Nature 354:82–84

    Article  CAS  PubMed  Google Scholar 

  3. Lam KS, Lebl M, Krchnák V (1997) The “one-bead-one-compound” combinatorial library method. Chem Rev 97:411–448

    Article  CAS  PubMed  Google Scholar 

  4. Yao N, Xiao W, Wang X, Marik J, Park SH, Takada Y, Lam KS (2009) Discovery of targeting ligands for breast cancer cells using the one-bead one-compound combinatorial method. J Med Chem 52:126–133

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  5. Aina OH, Marik J, Liu R, Lau DH, Lam KS (2005) Identification of novel targeting peptides for human ovarian cancer cells using "one-bead one-compound" combinatorial libraries. Mol Cancer Ther 4:806–813

    Article  CAS  PubMed  Google Scholar 

  6. Appell KC, Chung TDY, Solly KJ, Chelsky D (1998) Biological characterization of neurokinin antagonists discovered through screening of a combinatorial library. J Biomol Screen 3:19–27

    Article  CAS  Google Scholar 

  7. Lam KS, Liu R, Miyamoto S, Lehman L, Tuscano JM (2003) Applications of one-bead one-compound combinatorial libraries and chemical microarrays in signal transduction research. Acc Chem Res 36:370–377

    Article  CAS  PubMed  Google Scholar 

  8. Abato P, Conroy JL, Seto CT (1999) Combinatorial library of serine and cysteine protease inhibitors that interact with both the S and S′ binding sites. J Med Chem 42:4001–4009

    Article  CAS  PubMed  Google Scholar 

  9. St Hilaire PM, Alves LC, Herrera F, Renil M, Sanderson SJ, Mottram JC, Coombs GH, Juliano MA, Juliano L, Arevalo J, Meldal M (2002) Solid-phase library synthesis, screening, and selection of tight-binding reduced peptide bond inhibitors of a recombinant Leishmania mexicana cysteine protease B. J Med Chem 45:1971–1982

    Article  CAS  PubMed  Google Scholar 

  10. Mc Bride JD, Freeman N, Domingo GJ, Leatherbarrow RJ (1996) Selection of chymotrypsin inhibitors from a conformationally-constrained combinatorial peptide library. J Mol Biol 259:819–827

    Article  CAS  Google Scholar 

  11. Chirayil S, Chirayil R, Luebke J (2009) Discovering ligands for a microRNA precursor with peptoid microarrays. Nucleic Acids Res 37:5486–5497

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  12. Peng L, Liu R, Marik J, Wang X, Takada Y, Lam KS (2006) Combinatorial chemistry identifies high-affinity peptidomimetics against a4b1 integrin for in vivo tumor imaging. Nat Chem Biol 2:381–389

    Article  CAS  PubMed  Google Scholar 

  13. Maverakis E, Menezes JS, Ametani A, Han M, Stevens DB, He Y, Wang Y, Ono Y, Lam KS, Ward ED, Sercarz EE (2010) Molecular mimics can induce a nonautoaggressive repertoire that preempts induction of autoimmunity. Proc Natl Acad Sci 107:2550–2555

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  14. Townsend JB, Shaheen F, Liu R, Lam KS (2010) Jeffamine derivatized tentagel beads and poly(dimethylsiloxane) microbead cassettes for ultrahigh-throughput in situ releasable solution-phase cell-based screening of one-bead-one-compound combinatorial small molecule libraries. J Comb Chem 12:700–712

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  15. Marani MM, Ceron MCM, Giudicessi SL, Oliveira E, Côté S, Erra-Balsells R, Albericio F, Cascone O, Camperi SA (2009) Screening of one-bead-one-peptide combinatorial library using red fluorescent dyes. Presence of positive and false positive beads. J Comb Chem 11:146–150

    Article  CAS  PubMed  Google Scholar 

  16. Luo J, Zhang H, Xiao W, Kumaresan PR, Shi C, Pan C, Aina OH, Lam KS (2008) Rainbow beads: a color coding method to facilitate high-throughput screening and optimization of one-bead one-compound combinatorial libraries. J Comb Chem 10:599–604

    Article  CAS  PubMed  Google Scholar 

  17. Xiao W, Bononi FC, Townsend J, Li Y, Liu R, Lam KS (2013) Immobilized OBOC combinatorial bead array to facilitate multiplicative screening. Comb Chem High Throughput Screen 16:441–448

    Article  CAS  PubMed  Google Scholar 

  18. Cho CF, Amadei GA, Breadner D, Luyt LG, Lewis JD (2012) Discovery of novel integrin ligands from combinatorial libraries using a multiplex “beads on a bead” approach. Nano Lett 12:5957–5965

    Article  CAS  PubMed  Google Scholar 

  19. Cho CF, Behnam Azad B, Luyt LG, Lewis JD (2013) High-throughput screening of one-bead-one-compound peptide libraries using intact cells. ACS Comb Sci 15:393–400

    Article  CAS  PubMed  Google Scholar 

  20. Liu R, Lam KS (2001) Automatic Edman microsequencing of peptides containing multiple unnatural amino acids. Anal Biochem 295:9–16

    Article  CAS  PubMed  Google Scholar 

  21. Amadei GA, Cho C-F, Lewis JD, Luyt LG (2009) A fast, reproducible and low-cost method for sequence deconvolution of ‘on-bead’ peptides via ‘on-target’ maldi-TOF/TOF mass spectrometry. J Mass Spectrom 45:241–251

    Article  Google Scholar 

  22. Youngquist RS, Fuentes GR, Lacey MP, Keough Baillie TA (2005) Matrix-assisted laser desorption ionization for rapid determination of the sequences of biologically active peptides isolated from support-bound combinatorial peptide libraries. Rapid Comm Mass Spec 8:77–81

    Article  Google Scholar 

  23. Semmler A, Weber R, Przybylski M, Wittmann V (2010) De novo sequencing of peptides on single resin beads by MALDI-FTICR tandem mass spectrometry. J Am Soc Mass Spectrom 21:215–219

    Article  CAS  PubMed  Google Scholar 

  24. Chan WC, White PD (2000) Fmoc solid phase peptide synthesis: a practical approach. Oxford University Press, New York, NY

    Google Scholar 

  25. Steen H, Mann M (2004) The ABC’s (and XYZ’s) of peptide sequencing. Nat Rev Mol Cell Biol 5:699–711

    Article  CAS  PubMed  Google Scholar 

  26. Yergey AL, Coorssen JR, Backlund PS Jr, Blank PS, Humphrey GA, Zimmerberg J, Campbell JM, Vestal ML (2002) De novo sequencing of peptides using MALDI/TOF-TOF. J Am Soc Mass Spectrom 13(7):784–791

    Article  CAS  PubMed  Google Scholar 

  27. Kaiser E, Colescott RL, Bossinger CD, Cook PI (1970) Color test for detection of free terminal amino groups in the solid-phase synthesis of peptides. Anal Biochem 34:595–598

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the Natural Sciences and Engineering Research Council of Canada (NSERC) and the Ontario Institute for Cancer Research (OICR).

Author information

Authors and Affiliations

  1. Department of Oncology, The University of Western Ontario, London, ON, Canada

    Leonard G. Luyt

  2. Department of Medical Imaging, The University of Western Ontario, London, ON, Canada

    Leonard G. Luyt

  3. Department of Chemistry, The University of Western Ontario, London, ON, Canada

    Fernanda C. Bononi & Leonard G. Luyt

  4. Cancer Research Laboratory Program, London Regional Cancer Program, London Health Sciences Centre, Room A4-817A, 790 Commissioners Rd. E, London, ON, Canada, N6A 4L6

    Fernanda C. Bononi & Leonard G. Luyt

Authors
  1. Fernanda C. Bononi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Leonard G. Luyt
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Leonard G. Luyt .

Editor information

Editors and Affiliations

  1. Dept of Chemistry, University of Alberta, Edmonton, Alberta, Canada

    Ratmir Derda

Rights and permissions

Reprints and permissions

Copyright information

© 2015 Springer Science+Business Media New York

About this protocol

Cite this protocol

Bononi, F.C., Luyt, L.G. (2015). Synthesis and Cell-Based Screening of One-Bead-One-Compound Peptide Libraries. In: Derda, R. (eds) Peptide Libraries. Methods in Molecular Biology, vol 1248. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2020-4_15

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-2020-4_15

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2019-8

  • Online ISBN: 978-1-4939-2020-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation