Toward single molecule detection of staphylococcal enterotoxin B: mobile sandwich immunoassay on gliding microtubules.
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.
Soto
Carissa M
CM
Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, 4555 Overlook Avenue SW, Washington, DC 20375, USA. carissa.soto@nrl.navy.mil
Martin
Brett D
BD
Sapsford
Kim E
KE
Blum
Amy Szuchmacher
AS
Ratna
Banahalli R
BR
D004768:Enterotoxins / Q000032:analysis* / Q000276:immunology*; D007118:Immunoassay / Q000379:methods*; D008870:Microtubules; D013050:Spectrometry, Fluorescence
D004768:Enterotoxins; C031818:enterotoxin B, staphylococcal