Anti-rabbit immunoglobulin G detection in complex medium by PM-RAIRS and QCM Influence of the antibody immobilisation method
|Author(s)||Briand Elisabeth1, Salmain M2, Compere Chantal3, Pradier C1|
|Affiliation(s)||1 : Univ Paris 06, CNRS, UMR 7609, Lab React Surface, F-75252 Paris 05, France.
2 : Ecole Natl Super Chim Paris, UMR 7576, Lab Chim & Biochim Complexes Mol, F-75231 Paris, France.
3 : IFREMER, Ctr Brest, Dept Essais & Rech Technol Interfaces & Capteurs, F-29280 Plouzane, France.
|Source||Biosensors and Bioelectronics (0956-5663) (Elsevier), 2007-06 , Vol. 22 , N. 12 , P. 2884-2890|
|WOS© Times Cited||39|
|Keyword(s)||RIgG immobilisation, PM RAIRS, QCM, Immunosensors|
|Abstract||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 trticrobalance (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. (c) 2006 Elsevier B.V. All rights reserved.|