Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 1996 Apr;70(4):1985–1995. doi: 10.1016/S0006-3495(96)79763-1

Surface engineering: optimization of antigen presentation in self-assembled monolayers.

C Duschl 1, A F Sévin-Landais 1, H Vogel 1
PMCID: PMC1225167  PMID: 8785357

Abstract

The formation of self-assembled monolayers (SAMs) on gold surfaces containing an antigenic peptide (NANP)6 and HS(CH2)11OH, and the specific binding of a monoclonal antibody to these layers were investigated by surface plasmon resonance (SPR). Peptides were synthesized by solid-state phase synthesis and were linked either to cysteine or to an alkyl-thiol to allow covalent attachment to gold. The content of the peptide in the SAMs was systematically varied, and the binding properties of the monoclonal antibody were compared with those measured by microcalorimetry in solution. At a critical peptide concentration in the SAM an optimal antibody binding and complete surface coverage was attained. At lower peptide concentrations, the amount of adsorbed antibody decreased; at higher peptide concentrations, the binding constant decreased. These effects can be explained if the accessibility of the antigenic epitopes depends on the peptide density. Addition of free antigen induced the desorption of bound antibodies and allowed accurate measurements of the dissociation rate constant. Binding constants obtained from steady-state measurements and from measurements of the kinetic rate constants were compared.

Full text

PDF
1985

Images in this article

1986FIGURE 1
on p.1986

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amzel L. M., Poljak R. J. Three-dimensional structure of immunoglobulins. Annu Rev Biochem. 1979;48:961–997. doi: 10.1146/annurev.bi.48.070179.004525. [DOI] [PubMed] [Google Scholar]
  2. Berg H. C., Purcell E. M. Physics of chemoreception. Biophys J. 1977 Nov;20(2):193–219. doi: 10.1016/S0006-3495(77)85544-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bernard A., Bosshard H. R. Real-time monitoring of antigen-antibody recognition on a metal oxide surface by an optical grating coupler sensor. Eur J Biochem. 1995 Jun 1;230(2):416–423. doi: 10.1111/j.1432-1033.1995.0416h.x. [DOI] [PubMed] [Google Scholar]
  4. Fägerstam L. G., Frostell-Karlsson A., Karlsson R., Persson B., Rönnberg I. Biospecific interaction analysis using surface plasmon resonance detection applied to kinetic, binding site and concentration analysis. J Chromatogr. 1992 Apr 24;597(1-2):397–410. doi: 10.1016/0021-9673(92)80137-j. [DOI] [PubMed] [Google Scholar]
  5. Goldstein B., Posner R. G., Torney D. C., Erickson J., Holowka D., Baird B. Competition between solution and cell surface receptors for ligand. Dissociation of hapten bound to surface antibody in the presence of solution antibody. Biophys J. 1989 Nov;56(5):955–966. doi: 10.1016/S0006-3495(89)82741-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Karlsson R., Michaelsson A., Mattsson L. Kinetic analysis of monoclonal antibody-antigen interactions with a new biosensor based analytical system. J Immunol Methods. 1991 Dec 15;145(1-2):229–240. doi: 10.1016/0022-1759(91)90331-9. [DOI] [PubMed] [Google Scholar]
  7. Nussenzweig V., Nussenzweig R. S. Rationale for the development of an engineered sporozoite malaria vaccine. Adv Immunol. 1989;45:283–334. doi: 10.1016/s0065-2776(08)60695-1. [DOI] [PubMed] [Google Scholar]
  8. Ortega E., Schweitzer-Stenner R., Pecht I. Kinetics of ligand binding to the type 1 Fc epsilon receptor on mast cells. Biochemistry. 1991 Apr 9;30(14):3473–3483. doi: 10.1021/bi00228a018. [DOI] [PubMed] [Google Scholar]
  9. Panayotou G., Gish G., End P., Truong O., Gout I., Dhand R., Fry M. J., Hiles I., Pawson T., Waterfield M. D. Interactions between SH2 domains and tyrosine-phosphorylated platelet-derived growth factor beta-receptor sequences: analysis of kinetic parameters by a novel biosensor-based approach. Mol Cell Biol. 1993 Jun;13(6):3567–3576. doi: 10.1128/mcb.13.6.3567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Petrossian A., Owicki J. C. Interaction of antibodies with liposomes bearing fluorescent haptens. Biochim Biophys Acta. 1984 Oct 3;776(2):217–227. doi: 10.1016/0005-2736(84)90211-6. [DOI] [PubMed] [Google Scholar]
  11. Pisarchick M. L., Thompson N. L. Binding of a monoclonal antibody and its Fab fragment to supported phospholipid monolayers measured by total internal reflection fluorescence microscopy. Biophys J. 1990 Nov;58(5):1235–1249. doi: 10.1016/S0006-3495(90)82464-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Wiseman T., Williston S., Brandts J. F., Lin L. N. Rapid measurement of binding constants and heats of binding using a new titration calorimeter. Anal Biochem. 1989 May 15;179(1):131–137. doi: 10.1016/0003-2697(89)90213-3. [DOI] [PubMed] [Google Scholar]
  13. Wohlhueter R. M., Parekh K., Udhayakumar V., Fang S., Lal A. A. Analysis of binding of monoclonal antibody to a malarial peptide by surface plasmon resonance biosensor and integrated rate equations. J Immunol. 1994 Jul 1;153(1):181–189. [PubMed] [Google Scholar]
  14. van den Heuvel D. J., Kooyman R. P., Drijfhout J. W., Welling G. W. Synthetic peptides as receptors in affinity sensors: a feasibility study. Anal Biochem. 1993 Dec;215(2):223–230. doi: 10.1006/abio.1993.1579. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES