Clickable Polymeric Coating for Oriented Peptide Immobilization

  • Laura SolaEmail author
  • Alessandro Gori
  • Marina Cretich
  • Chiara Finetti
  • Caterina Zilio
  • Marcella Chiari
Part of the Methods in Molecular Biology book series (MIMB, volume 1352)


A new methodology for the fabrication of an high-performance peptide microarray is reported, combining the higher sensitivity of a layered Si–SiO2 substrate with the oriented immobilization of peptides using a N,N-dimethylacrylamide-based polymeric coating that contains alkyne monomers as functional groups. This clickable polymer allows the oriented attachment of azido-modified peptides via a copper-mediated azide/alkyne cycloaddition. A similar coating that does not contain the alkyne functionality has been used as comparison, to demonstrate the importance of a proper orientation for facilitating the probe recognition and interaction with the target antibody.

Key words

Peptide microarray Click chemistry Oriented immobilization Polymeric coating 


  1. 1.
    Fu J, Reinhold J, Woodbury NW (2011) Peptide-modified surfaces for enzyme immobilization. PLoS One 6:e18692CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Rao SV, Anderson KW, Bachas LG (1998) Oriented immobilization of proteins. Microkim Acta 128:127–143CrossRefGoogle Scholar
  3. 3.
    Van Berkel SS, Van Eldijk MB, Van Hest JCM (2011) Staudinger ligation as a method for bioconjugation. Angew Chem Int Ed 50:8806–8827CrossRefGoogle Scholar
  4. 4.
    Jung Y, Min Lee J, Kim J, Yoon J, Cho H, Chung BH (2009) Photoactivable antibody binding protein: site-selective and covalent coupling of antibody. Anal Chem 81:936–942CrossRefPubMedGoogle Scholar
  5. 5.
    Liang L, Astruc D (2011) The copper(I)-catalyzed alkyne-azide cycloaddition (CuAAC) “click” reaction and its applications. An overview. Coordin Chem Rev 255:2933–2945CrossRefGoogle Scholar
  6. 6.
    Huisgen R (1989) Kinetics and reaction mechanisms: selected examples from the experience of forty years. Pure Appl Chem 61:613–628CrossRefGoogle Scholar
  7. 7.
    Pirri G, Damin F, Chiari M, Bontempi E, Depero LE (2004) Characterization of a polymeric adsorbed coating for DNA microarray glass slides. Anal Chem 76:1352–1358CrossRefPubMedGoogle Scholar
  8. 8.
    Zilio C, Bernardi A, Palmioli A, Salina M, Tagliabue G, Buscaglia M, Consonni R, Chiari M (2015) New “clickable” polymeric coating for glycan microarrays. Sensor Actuator B 215: 412–420Google Scholar
  9. 9.
    Uttamapinant C, Tangpeerachaikul A, Grecian S, Clarke S, Singh U, Slade P, Gee KR, Ting AY (2012) Fast, cell-compatible click chemistry with copper-chelating azides for biomolecular labeling. Angew Chem Int Ed 51:5852–5856CrossRefGoogle Scholar
  10. 10.
    Peri C, Gagni P, Combi F, Gori A, Chiari M, Longhi R, Cretich M, Colombo G (2013) Rational epitope design for protein targeting. ACS Chem Biol 8:397–404CrossRefPubMedGoogle Scholar
  11. 11.
    Cretich M, Monroe MR, Reddington A, Zhang X, Daaboul GG, Damin F, Sola L, Unlu SM, Chiari M (2012) Interferometric silicon biochips for label and label-free DNA and protein microarrays. Proteomics 12:2963–2977CrossRefPubMedGoogle Scholar
  12. 12.
    Ladmiral V, Mantovani G, Clarkson GJ, Cauet S, Irwin JL, Haddleton DM (2006) Synthesis of neoglycopolymers by a combination of “click chemistry” and living radical polymerization. J Am Chem Soc 128:4823–4830CrossRefPubMedGoogle Scholar
  13. 13.
    Sola L, Chiari M (2012) Modulation of electroosmotic flow in capillary electrophoresis using functional polymer coatings. J Chrom A 1270:324–329CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Laura Sola
    • 1
    Email author
  • Alessandro Gori
    • 1
  • Marina Cretich
    • 1
  • Chiara Finetti
    • 1
  • Caterina Zilio
    • 1
  • Marcella Chiari
    • 1
  1. 1.Istituto di Chimica del Riconoscimento Molecolare (ICRM), Consiglio Nazionale delle Ricerche (CNR)MilanItaly

Personalised recommendations