Gas phase activity of anti-FITC antibodies immobilized on a surface acoustic wave resonator device.
In this paper we present the results of a series of experiments on the activity of antibodies in a vapor phase sensor. For these experiments the sensor component was a ST-Quartz resonator with a center frequency of approximately 250 MHz. Anti-FITC antibodies were attached to the electrodes on the device surface via a protein-A crosslinker. Surface acoustic wave (SAW) resonator devices with various coatings were mounted in TO-8 packages, inserted into our sensor head module and subjected to various fluorescent analyte gases. Numerous controls were performed including the use of coated and uncoated devices along with devices coated with antibodies which were not specific for the target analyte. The SAW immunosensor response was monitored and a baseline frequency shift was observed when the analyte being presented was the antigen for the immobilized antibody. To provide an independent measure of antibody/antigen binding, the devices were removed from the sensor head, washed with a buffer solution to remove any unbound analyte, and then inspected using a confocal laser scanning microscope (CLSM). Since all the analytes being used in these experiments were fluorescent this afforded us the opportunity to visualize the attachment of the analyte to the antibody film. Given the high resolution of the CLSM, we were able to identify the location of the attachment of the fluorescent analytes relative to the 1.5 microm wide electrodes of the SAW device. We believe that these experiments demonstrate that we have achieved real time molecular recognition of these small molecules in the vapor phase.
Stubbs
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School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA 30332, USA.
Hunt
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Lee
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Doyle
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D000162:Acoustics / Q000295:instrumentation; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000032:analysis; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000936:Antigen-Antibody Complex / Q000032:analysis* / Q000378:metabolism; D000941:Antigens / Q000032:analysis* / Q000378:metabolism; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D004867:Equipment Design; D005240:Feasibility Studies; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry / Q000378:metabolism*; D005455:Fluorescent Antibody Technique / Q000295:instrumentation / Q000379:methods; D005740:Gases / Q000032:analysis*; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D008958:Models, Molecular; D008962:Models, Theoretical; D012680:Sensitivity and Specificity; D013205:Staphylococcal Protein A / Q000378:metabolism; D013499:Surface Properties
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000941:Antigens; D005740:Gases; D013205:Staphylococcal Protein A; D016650:Fluorescein-5-isothiocyanate