Localization and quantitative analysis of antigen-antibody binding on 2D substrate using imaging NanoSIMS |
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Authors: | Dauphas Stéphanie Delhaye Thomas Lavastre Olivier Corlu Anne Guguen-Guillouzo Christiane Ababou-Girard Soraya Geneste Florence |
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Affiliation: | Laboratoire des Sciences Chimiques de Rennes, Equipe catalyse et organométalliques, Université de Rennes 1, UMR-CNRS 6226, Campus de Beaulieu, 35042 Rennes cedex, France. |
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Abstract: | Iron containing-antigen bound specifically to antibody immobilized on a surface is analyzed by nanoscale secondary ion mass spectrometry (NanoSIMS). This technique is well adapted compared with X-ray photoelectron spectroscopy and energy dispersive spectroscopy, which do not allow the detection of iron. The obtained Fe(+) map gives a good representation of the antigen repartition on the surface. NanoSIMS analysis of competition experiments performed with albumin and iron-free antigen are in good accordance with results obtained by a classical fluorescence microscopy approach. These results underline the interest of imaging NanoSIMS as a label-free method, allowing the localization and quantitative analysis of antigen-antibody binding with better spatial resolution than imaging ellipsometry and SPR. |
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