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    Anti-rabbit immunoglobulin G detection in complex medium by PM-RAIRS and QCM: Influence of the antibody immobilisation method

    Year: 2007

    Journal: Biosensors and Bioelectronics, Volume 22, Issue 12, 15 June 2007, Pages 2884-2890, 20111221

    Authors: Elisabeth Briand, Michèle Salmain, Chantal Compère and Claire-Marie Pradier

    Organizations: Laboratoire de Réactivité de Surface, UMR CNRS 7609, Université Pierre et Marie Curie, 4 place Jussieu, 75252 Paris Cedex 05, France, Laboratoire de Chimie et Biochimie des Complexes Moléculaires, UMR 7576, Ecole Nationale Supérieure de Chimie Paris, 75231 Paris Cedex 05, France, Département Essais et Recherches Technologiques, Interfaces et capteurs, IFREMER, Centre de Brest, BP 70, 29280 Plouzané, France

    Two antibody immobilisation procedures were compared to set up an immunosensor for goat anti-rabbit immunoglobulin (anti-rIgG), i.e. rIgG covalently bound or immobilised via affinity to protein A (PrA). In both cases, the first layer of protein was covalently bound to a mixed self-assembled monolayer (SAM) of mercaptoundecanoic acid (MUA) and mercaptohexanol (C6OH) on a gold surface. The elaboration of the sensitive surfaces, as well as their selectivity and sensitivity were studied step by step by polarization modulation-reflection absorption infra-red spectroscopy (PM-RAIRS) and quartz crystal microbalance (QCM) with impedance measurement. QCM measurements showed that the viscoelastic properties of the antibody layer were markedly modified during the antigen recognition when the antibody was bound by affinity to PrA. The specific detection of antigen within a complex medium was assessed by PM-RAIRS thanks to the grafting of cobalt-carbonyl probes. Affinity constants between the immobilised rIgG and the anti-rIgG were determined from PM-RAIRS analysis.