Orientation of adsorbed antibodies: in situ monitoring by QCM and random sequential adsorption modeling.
(2012) Proteins at Interfaces III: State of the Art. — ISBN: [ISBN13: 9780841227965], p. 453-469, published
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- Antibodies (IgGs) are widely used for diagnostic assays, for which they are in certain cases immobilized by adsorption on hydrophobic substrates. Antigen recognition efficiency will depend on the orientation of the adsorbed IgG molecules. The aim of the present study was to investigate the binding-ability of a range of IgG isotypes from rat and mouse, all directed against the same antigen, using quartz crystal microbalance. The results allow identifying some isotypes which adsorb in higher amount and which provide a better bound antigen to adsorbed IgG ratio. This ratio was found to remain rather constant with the adsorbed IgG amount. Random sequential adsorption (RSA) modeling was used to simulate IgG adsorption. In the chosen modeling conditions, it is shown that even if adsorption in flat orientation is more favorable, a high proportion of IgG molecules adsorb in end-on orientation when surface coverage increases, owing to the low surface area spaces left between IgG molecules already adsorbed in flat orientation. The apparent discrepancy between experimental data collected by QCM and the output of RSA modeling may be attributed to variations in the water content of the adsorbed layer, to steric hindrance and multivalency effects upon antigen binding, or to the role of albumin molecules used to prevent non specific adsorption of the antigen.
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Dupont, C. (2012). Orientation of adsorbed antibodies: in situ monitoring by QCM and random sequential adsorption modeling. In Horbett, T., et al. (ed.), Proteins at Interfaces III: State of the Art. (p. p. 453-469). American Chemical Society. https://doi.org/10.1021/bk-2012-1120