Array biosensor for detection of toxins

Abstract

The array biosensor is capable of detecting multiple targets rapidly and simultaneously on the surface of a single waveguide. Sandwich and competitive fluoroimmunoassays have been developed to detect high and low molecular weight toxins, respectively, in complex samples. Recognition molecules (usually antibodies) were first immobilized in specific locations on the waveguide and the resultant patterned array was used to interrogate up to 12 different samples for the presence of multiple different analytes. Upon binding of a fluorescent analyte or fluorescent immunocomplex, the pattern of fluorescent spots was detected using a CCD camera. Automated image analysis was used to determine a mean fluorescence value for each assay spot and to subtract the local background signal. The location of the spot and its mean fluorescence value were used to determine the toxin identity and concentration. Toxins were measured in clinical fluids, environmental samples and foods, with minimal sample preparation. Results are shown for rapid analyses of staphylococcal enterotoxin B, ricin, cholera toxin, botulinum toxoids, trinitrotoluene, and the mycotoxin fumonisin. Toxins were detected at levels as low as 0.5 ng mL(-1).

Sensing the Deadliest Toxin: Technologies for Botulinum Neurotoxin Detection

“…Using this system, multiple biosensors were tested with BoNTs [98,99,100,101,102,103]. Sensitivities of the sensors range from 150 pg/mL (15 MLD 50 /mL) for BoNT/E [100] to 200 ng/mL for BoNT/B [101] in buffers.…”

“…Sensitivities of the sensors range from 150 pg/mL (15 MLD 50 /mL) for BoNT/E [100] to 200 ng/mL for BoNT/B [101] in buffers. Besides single analyte sensors, array sensors for multiple analytes were constructed for the simultaneous detection of several toxins such as BoNT/A, /B, cholera, ricin and staphylococcal endotoxin B, and bacterial samples [101,102]. Detection of BoNT/A toxoid in various food samples has been reported as well with sensitivities typically around 50 ng/mL [103].…”

Interfacing "Soft" and "Hard" Matter with Exquisite Chemical Control

“…Protein chips can also be used as a diagnosis tool for profiling protein expression in order to find potentially relevant biomarkers [67]. Another potential application for ordered protein films on surfaces will be the production of optimized biosensors [68,69]. An ordered protein film has a higher activity density than a random protein film, where a significant percentage of the protein molecules bound to the surface are potentially inactive conformations.…”

Biosensor Arrays for Environmental Monitoring

“…Biosensor array that is capable of detecting multiple targets rapidly and simultaneously on the surface of a single waveguide has also been studied. Ligler et al [31] developed a sandwich and competitive fluoroimmunoassays to detect high and low molecular weight toxins, respectively, in complex samples. Antibodies were first immobilized in specific locations on the waveguide and the resultant patterned array was used to interrogate up to 12 different samples for the presence of multiple different analytes.…”