In-depth investigation of protein adsorption on gold surfaces: correlating the structure and density to the efficiency of the sensing layer - INRAE - Institut national de recherche pour l’agriculture, l’alimentation et l’environnement Accéder directement au contenu
Article Dans Une Revue Journal of Physical Chemistry B Année : 2008

In-depth investigation of protein adsorption on gold surfaces: correlating the structure and density to the efficiency of the sensing layer

Résumé

Protein A (PrA), mouse monoclonal anti-IgG antibody (SAb) and deglycosylated avidin (NAV) were adsorbed on gold surfaces to capture the model rabbit IgG and build three immunosensing platforms. The assembling of immunosensors, their specificity, and the receptor accessibility were monitored by polarization modulation reflection-absorption infrared spectroscopy (PM-RAIRS) and quartz crystal microbalance with dissipation measurement (QCM-D) at each step. Combining these two techniques allows us to compare both chemical and structural properties of the sensing layers with the former bringing chemical and semiquantitative information on the grafted protein layers, whereas the latter, in addition to the mass uptake, enables us to take the layer rigidity into account. Grafting of the three capture proteins to the transducer surfaces, covered with appropriate self-assembled monolayers, yielded protein layers with variable properties. NAV formed a dense and rigid molecular layer, likely containing protein aggregates, whereas the amount of PrA was below one monolayer resulting in a flexible layer. The amount of immobilized rabbit IgG was different for the three systems with the densest capture protein layer exhibiting the lowest binding capacity. The accessibility of antibodies on the resulting immunosensors measured by interaction with a secondary antirabbit IgG antibody was found to be closely dependent on their coverage as well as on the rigidity of the protein layer. The overall study provides in-depth information on three of the most common immunosensor recognition interfaces and demonstrates the crucial influence of both structure and density of the protein layer on the efficiency of the molecular recognition phenomena.

Dates et versions

hal-02663385 , version 1 (31-05-2020)

Identifiants

Citer

Souhir Boujday, Aurore Bantegnie, Elisabeth Briand, Pierre-Guy Marnet, Michèle Salmain, et al.. In-depth investigation of protein adsorption on gold surfaces: correlating the structure and density to the efficiency of the sensing layer. Journal of Physical Chemistry B, 2008, 112 (21), pp.6708-6715. ⟨10.1021/jp711916g⟩. ⟨hal-02663385⟩
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