Study of immunoglobulin G thin layers obtained by the Langmuir-Blodgett method: application to immunosensors.
Nowadays, immunosensors play a leading part in the field of bioanalytical chemistry research. As with any biosensor, they need appropriate transducers and a suitable technique to immobilize the active biocomponents. In this study, two transduction modes were chosen: mass effects (quartz microbalance measurements) and geometric and dielectric effects (capacitance measurements). The Langmuir-Blodgett (LB) method appears to be quite suitable for generating biospecific surfaces. This work has focused on the detection of staphylococcal enterotoxin B, the corresponding antibody being immobilized at the surface of fatty acids by a variant of the LB method. The composition of the film and the nature of antibody-fatty acid interactions were studied by means of the two transducers mentioned above. FTIR (Fourier transform infra-red) spectroscopy and protein diagnostic assay. Influence of several parameters (pH, ionic strength, transfer pressure, antibody concentration in the subphase) was investigated. The immobilization rate reached its maximum when experimental conditions allowed optimal electrostatic interactions. In this case, the quartz crystal microbalance response, in air, reached 55 Hz per monolayer of immobilized immunoglobulin G and the equivalent capacitance variation, measured in liquid media, was around 300 pF cm-2. Activity of the biospecific LB films, when binding enterotoxin, was checked by the classical ELISA (enzyme immuno-linked assay) technique.
Barraud
A
A
Centre d'Etudes Nucléaires de Saclay, DRECAM, SCM, Gif-sur-Yvette, France.
Perrot
H
H
Billard
V
V
Martelet
C
C
Therasse
J
J
D000937:Antigen-Antibody Reactions; D015374:Biosensing Techniques; D004768:Enterotoxins / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D005227:Fatty Acids; D005583:Fourier Analysis; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G; D008722:Methods; D009994:Osmolar Concentration; D013055:Spectrophotometry, Infrared / Q000379:methods; D013211:Staphylococcus aureus; D014159:Transducers
D004768:Enterotoxins; D005227:Fatty Acids; D007074:Immunoglobulin G; C031818:enterotoxin B, staphylococcal