Ultrasensitive evanescent wave fluoro-immunosensors using polystyrene integrated lens optical fiber

A sensitive evanescent wave immunoassay is described for assay of human chorionic gonadotropin (hCG) in plasma using molded polystyrene optical fibers. Data was processed using kinetic parameters which allowed a total assay time of two minutes. The characterization of two longer wavelength dyes (allophycocyanin and Cy5) in relation to FITC is described and their effect on assay signal was studied. APC had six times the fluorescence intensity of FITC and generated forty times the signal in the hCG assay format. Native, acid-treated and biotinylated antibody immobilization chemistries and their surface density effects were investigated to determine the effect on assay signal. Native antibody generated approximately 50% of the signal versus acid-treated or biotinylated and a 50% reduction in the surface antibody concentration showed little effect on assay signal. The assay agreed well with a commercially available assay (ES-300) and has a sensitivity of 0.8 mIU/ml.

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