Skip to main content

Journal for Biophysical Chemistry

Biointerphases Cover Image

Site-specific presentation of single recombinant proteins in defined nanoarrays

Abstract

The authors describe the deposition of single biomolecules on substrates at defined spacing by pure self-assembly. The substrate is equipped with an array of 8 nm large gold particles which form the template for biomolecule binding. The authors verified the successful binding of single biomolecules via specific antibody labeling and imaging by fluorescence microscopy. Scanning force microscopy provided evidence that every gold nanoparticle of the pattern is occupied by at least one biomolecule. Furthermore, gold conjugated secondary antibodies in combination with scanning electron microscopy proved that at least 75% of the nanoparticles carried only one active biomolecule. The precision given by such surface densities is molecularly defined and such considerably higher than in any other case reported so far.

References

  1. 1

    D. L. Spector, R. D. Goldman, and L. A. Leinwand, Cells: A Laboratory Manual (CSHL, 1998).

  2. 2

    L. L. Kiessling, J. E. Gestwicki, and L. E. Strong, Angew. Chem., Int. Ed. 45, 2348 (2006).

    Article  CAS  Google Scholar 

  3. 3

    M. Arnold, E. A. Cavalcanti-Adam, R. Glass, J. Blummel, W. Eck, M. Kantlehner, H. Kessler, and J. P. Spatz, ChemPhysChem 19, 383 (2004).

    Article  Google Scholar 

  4. 4

    E. A. Cavalcanti-Adam, A. Micoulet, J. Blummel, J. Auernheimer, H. Kessler, and J. P. Spatz, Eur. J. Cell Biol. 85, 219 (2006).

    Article  CAS  Google Scholar 

  5. 5

    M. A. Cooper, Nat. Rev. Drug Discovery 1, 515 (2002).

    Article  CAS  Google Scholar 

  6. 6

    N. Green, Avidin and Streptavidin, Avidin-Biotin Technology, Methods in Enzymology, (Academic, San Diego, 1990).

    Google Scholar 

  7. 7

    J. Groll, K. Albrecht, P. Gasteier, S. Riethmueller, U. Ziener, and M. Moeller, ChemBioChem 6, 1782 (2005).

    Article  CAS  Google Scholar 

  8. 8

    G. Lemercier and K. Johnsson, Nat. Methods 3, 247 (2006).

    Article  CAS  Google Scholar 

  9. 9

    G. B. Sigal, C. Bamdad, A. Barberis, J. Strominger, and G. M. Whitesides, Anal. Chem. 68, 490 (1996).

    Article  CAS  Google Scholar 

  10. 10

    K. Ataka, F. Giess, W. Knoll, R. Naumann, S. Haber-Pohlmeier, B. Richter, and J. Heberle, J. Am. Chem. Soc. 126, 16199 (2004).

    Article  CAS  Google Scholar 

  11. 11

    L. Nieba et al., Anal. Biochem. 252, 217 (1997).

    Article  CAS  Google Scholar 

  12. 12

    J. Schmitt, H. Hess, and H. G. Stunnenberg, Mol. Biol. Rep. 18, 223 (1993).

    Article  CAS  Google Scholar 

  13. 13

    P. Hinterdorfer and Y. F. Dufrene, Nat. Methods 3, 347 (2006).

    Article  CAS  Google Scholar 

  14. 14

    S. Lata, M. Gavutis, R. Tampe, and J. Piehler, J. Am. Chem. Soc. 128, 2365 (2006).

    Article  CAS  Google Scholar 

  15. 15

    D. L. Johnson and L. L. Martin, J. Am. Chem. Soc. 127, 2018 (2005).

    Article  CAS  Google Scholar 

  16. 16

    V. N. Truskett and M. P. Watts, Trends Biotechnol. 24, 312 (2006).

    Article  CAS  Google Scholar 

  17. 17

    M. L. Dustin and D. R. Colman, Science 298, 785 (2002).

    Article  CAS  Google Scholar 

  18. 18

    B. F. Lillemeier, J. R. Pfeiffer, Z. Surviladze, B. S. Wilson, and M. M. Davis, Proc. Natl. Acad. Sci. U.S.A. 103, 18992 (2006).

    Article  CAS  Google Scholar 

  19. 19

    G. A. Silva, Nat. Rev. Neurosci. 7, 65 (2006).

    Article  CAS  Google Scholar 

  20. 20

    K. Busch and R. Tampe, Mol. Biotechnol. 82, 3 (2001).

    Article  CAS  Google Scholar 

  21. 21

    J. Spatz et al., Langmuir 16, 407 (2000).

    Article  CAS  Google Scholar 

  22. 22

    R. Glass, M. Moeller, and J. P. Spatz, Nanotechnology 14, 1153 (2003).

    Article  CAS  Google Scholar 

  23. 23

    G. Kadmon and P. Altevogt, Differentiation 61, 143 (1997).

    Article  CAS  Google Scholar 

  24. 24

    G. Tsen, W. Halfter, S. Kroger, and G. J. Cole, J. Biol. Chem. 270, 3392 (1995).

    Article  CAS  Google Scholar 

  25. 25

    M. Takeichi, Curr. Opin. Cell Biol. 7, 619 (1995).

    Article  Google Scholar 

  26. 26

    U. Tepass, K. Truong, D. Godt, M. Ikura, and M. Peifer, Nat. Rev. Mol. Cell Biol. 1, 91 (2000).

    Article  CAS  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Joachim P. Spatz.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Wolfram, T., Belz, F., Schoen, T. et al. Site-specific presentation of single recombinant proteins in defined nanoarrays. Biointerphases 2, 44–48 (2007). https://doi.org/10.1116/1.2713991

Download citation