Localization and quantitative analysis of antigen-antibody binding on 2D substrate using imaging NanoSIMS.
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.
Dauphas
Stéphanie
S
Laboratoire des Sciences Chimiques de Rennes, Equipe catalyse et organométalliques, Université de Rennes 1, UMR-CNRS 6226, Campus de Beaulieu, 35042 Rennes cedex, France.
Delhaye
Thomas
T
Lavastre
Olivier
O
Corlu
Anne
A
Guguen-Guillouzo
Christiane
C
Ababou-Girard
Soraya
S
Geneste
Florence
F
D000937:Antigen-Antibody Reactions; D006801:Humans; D033862:Iron-Binding Proteins / Q000032:analysis*; D036103:Nanotechnology / Q000295:instrumentation*; D012709:Serum Albumin / Q000032:analysis; D018629:Spectrometry, Mass, Secondary Ion / Q000295:instrumentation / Q000379:methods*; D014168:Transferrin / Q000032:analysis
D033862:Iron-Binding Proteins; D012709:Serum Albumin; D014168:Transferrin