JOURNAL ARTICLE

Toward single molecule detection of staphylococcal enterotoxin B: mobile sandwich immunoassay on gliding microtubules

Carissa M Soto, Brett D Martin, Kim E Sapsford, Amy Szuchmacher Blum, Banahalli R Ratna
Analytical Chemistry 2008 July 15, 80 (14): 5433-40
18543949
An immunoassay based on gliding microtubules (MTs) is described for the detection of staphylococcal enterotoxin B. Detection is performed in a sandwich immunoassay format. Gliding microtubules carry the antigen-specific "capture" antibody, and bound analyte is detected using a fluorescent viral scaffold as the tracer. A detailed modification scheme for the MTs postpolymerization is described along with corresponding quantification by fluorescence spectroscopy. The resultant antibody-MTs maintain their morphology and gliding capabilities. We report a limit of detection down to 0.5 ng/mL during active transport in a 30 min assay time and down to 1 ng/mL on static surfaces. This study demonstrates the kinesin/MT-mediated capture, transport, and detection of the biowarfare agent SEB in a microfluidic format.

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