Lab-On-a-Chip for carbon nanotubes based immunoassay detection of Staphylococcal Enterotoxin B (SEB).
We describe a new eight channel Lab-On-a-Chip (LOC) for a Carbon Nanotube (CNT) based immunoassay with optical detection of Staphylococcal Enterotoxin B (SEB) for food safety applications. In this work, we combined four biosensing elements: (1) CNT technology for primary antibody immobilization, (2) Enhanced Chemiluminescence (ECL) for light signal generation, (3) a cooled charge-coupled device (CCD) for detection and (4) polymer lamination technology for developing a point of care immunological assay for SEB detection. Our concept for developing versatile LOCs, which can be used for many different applications, is to use a modular design with interchangeable recognition elements (e.g. various antibodies) to determine the specificity. Polymer lamination technology was used for the fabrication of a six layer, syringe operated LOC capable of analyzing eight samples simultaneously. An anti-SEB antibody-nanotube mixture was immobilized onto a polycarbonate strip, to serve as an interchangeable ligand surface that was then bonded onto the LOC. SEB samples are loaded into the device and detected by an ELISA assay using Horse Radish Peroxidase (HRP) conjugated anti-SEB IgG as a secondary antibody and ECL, with detection by a previously described portable cooled CCD detector. Eight samples of SEB in buffer or soy milk were assayed simultaneously with a limit of detection of 0.1 ng mL(-1). CNT immobilization of the antibody increased the sensitivity of detection six fold. Use of a simple interchangeable immunological surface allows this LOC to be adapted to any immunoassay by simply replacing the ligand surface. A syringe was used to move fluids for this assay so no power is needed to operate the device. Our versatile portable point-of-care CCD detector combined with the LOC immunoassay method described here can be used to reduce the exposure of users to toxins and other biohazards when working outside the lab, as well as to simplify and increase sensitivity for many other types of immunological diagnostics and detection assays.
Yang
Minghui
M
Center for Advanced Sensor Technology, University of Maryland Baltimore County, MD 21250, USA.
Sun
Steven
S
Kostov
Yordan
Y
Rasooly
Avraham
A
D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods; D002417:Cattle; D015169:Colony Count, Microbial / Q000295:instrumentation / Q000379:methods; D004768:Enterotoxins / Q000032:analysis* / Q000276:immunology; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005504:Food Analysis / Q000295:instrumentation* / Q000379:methods; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D007118:Immunoassay / Q000295:instrumentation*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D008892:Milk / Q000737:chemistry / Q000382:microbiology*; D037742:Nanotubes, Carbon / Q000737:chemistry* / Q000648:ultrastructure; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D004768:Enterotoxins; D037742:Nanotubes, Carbon; C031818:enterotoxin B, staphylococcal