Voltage-induced inhibition of antigen-antibody binding at conducting optical waveguides.
Optical waveguides coated with electrically conducting indium-tin oxide (ITO) are demonstrated here as a new class of substrate for fluorescent immunosensors. These waveguides combine electrochemical control with evanescent excitation and image-based detection. Presented here are preliminary results utilizing these waveguides that demonstrate influence of waveguide voltage on antigen binding. Specifically, waveguide surfaces were bisected into electrically addressable halves, anti-ovalbumin immobilized in patterns on their surfaces, and a 1.3 V bias applied between waveguide halves in the presence of Cy5-labeled ovalbumin in 10 mM phosphate buffer (pH 7.4) containing 150 mM NaCl and 0.05% Tween-20. Fluorescence imaging indicated that binding of the antigen to positively biased waveguide halves was inhibited nearly 10-fold compared with negatively biased waveguide halves and unbiased controls. Furthermore, it is shown that ovalbumin binding to positively biased waveguide regions is regenerated after removal of applied voltage. These results suggest that electrochemical control of immunosensor substrates can be used as a possible strategy toward minimizing cross-reactive binding and/or nonspecific adsorption, immunosensor regeneration, and controlled binding.
Liron
Zvi
Z
George Mason University, Arlington, VA, USA.
Tender
Leonard M
LM
Golden
Joel P
JP
Ligler
Frances S
FS
D000818:Animals; D000936:Antigen-Antibody Complex / Q000737:chemistry*; D000937:Antigen-Antibody Reactions; D001666:Binding Sites, Antibody; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D004553:Electric Conductivity; D004563:Electrochemistry / Q000379:methods*; D004574:Electromagnetic Fields; D004867:Equipment Design; D016231:Fluorescence Polarization Immunoassay / Q000295:instrumentation* / Q000379:methods*; D005455:Fluorescent Antibody Technique / Q000295:instrumentation / Q000379:methods; D015200:Fluoroimmunoassay / Q000295:instrumentation / Q000379:methods; D005898:Glass; D008832:Microchemistry / Q000379:methods; D055095:Optics and Photonics; D010047:Ovalbumin / Q000032:analysis*; D011817:Rabbits; D012680:Sensitivity and Specificity; D017971:Tin Compounds / Q000737:chemistry*
D000936:Antigen-Antibody Complex; D020099:Coated Materials, Biocompatible; D017971:Tin Compounds; C109984:indium tin oxide; D010047:Ovalbumin