Functional molecular thin films: topological templates for the chemoselective ligation of antigenic peptides to self-assembled monolayers

Hans Jörg Mathieu, Horst Vogel
1999
Article

Résumé

Chem. ligation methods in combination with regioselective addressable templates have allowed the prepn. of well defined complex monomol. films tethered to gold surfaces. The supramol. monolayers exposed artificial recognition sites, here for monoclonal antibodies. SPR, FTIR spectroscopies, and TOF-SIMS were ideally suited surface sensitive techniques to enable the essential control of the individual reaction steps. The concept presented here for the functionalization of gold surfaces may be easily applied to other supporting materials, and is not limited to peptides, but may equally extend to proteins or nucleic acids. This approach may be extended to the construction of multifunctional surfaces with potential in bio-analytics. [on SciFinder (R)]

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https://infoscience.epfl.ch/record/81068?ln=fr

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Publications associées (9)

Direct Observation of Self-Assembled Monolayers, Ion Complexation, and Protein Conformation at the Gold/Water Interface: An FTIR Spectroscopic Approach

Claus Duschl, Martha Liley, Horst Vogel

A simple approach to the in situ measurement of IR spectra of self-assembled monolayers (SAMs) on gold in an aq. environment is described. A thin gold film (3-5 nm) on the surface of a germanium internal reflection element is used as a substrate for the self-assembly of a monolayer of a chelating thioalkane. High signal-to-noise spectra are obtained, allowing identification of the chem. groups present in the SAM. Complexation of ions by the SAM can be monitored, and the secondary conformation of His-tagged proteins bound to the SAM can be obsd. [on SciFinder (R)]

1997

Chargement

The formation of self-assembled monolayers (SAMs) on gold surfaces contg. 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 soln. At a crit. peptide concn. in the SAM an optimal antibody binding and complete surface coverage was attained. At lower peptide concns., the amt. of adsorbed antibody decreased: at higher peptide concns., the binding const. decreased. These effects can be explained if the accessibility of the antigenic epitopes depends on the peptide d. Addn. of free antigen induced the desorption of bound antibodies and allowed accurate measurements of the dissocn. rate const. Binding consts. obtained from steady-state measurements and from measurements of the kinetic rate consts. were compared. [on SciFinder (R)]

1996

Chargement

Reversible immobilization of peptides: Surface modification and in situ detection by attenuated total reflection FTIR spectroscopy

Patrik Hoffmann, Michael Mayer, Per Rigler, Horst Vogel

A generic method is described for the reversible immobilization of polyhistidine-bearing polypeptides and proteins on attenuated total reflecting (ATR) sensor surfaces for the detection of biomol. interactions by FTIR spectroscopy. Nitrilotriacetic acid (NTA) groups are covalently attached to self-assembled monolayers of either thioalkanes on gold films or mercaptosilanes on silicon dioxide films deposited on germanium internal reflection elements. Complex formation between Ni2+ ions and NTA groups activates the ATR sensor surface for the selective binding of polyhistidine sequences. This approach not only allows a stable and reversible immobilization of histidine-tagged peptides (His-peptides) but also simultaneously allows the direct in situ quantification of surface-adsorbed mols. from their specific FTIR spectral bands. The surface concns. of both NTA and His-peptide on silanized surfaces were detd. to be 1.1 and 0.4 mols. nm-2, resp., which means that the surface is densely covered. A comparison of exptl. FTIR spectra with simulated spectra reveals a surface-enhancement effect of one order of magnitude for the gold surfaces. With the presented sensor surfaces, new ways are opened up to investigate, in situ and with high sensitivity and reproducibility, protein-ligand, protein-protein, protein-DNA interactions, and DNA hybridization by ATR-FTIR spectroscopy. [on SciFinder (R)]

2003

Chargement

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