Laser-Induced Fluorescence Detector for Capillary-Based Isoelectric Immunoblot Assay

We describe a whole-capillary, multicolor laser-induced fluorescence scanner for microfluidic protein analysis systems. Separation of proteins is achieved by isoelectric focusing in a short length of fused-silica capillary after which the resolved proteins are immobilized to the capillary wall using...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2007-12, Vol.79 (24), p.9478-9483
Main Authors: Knittle, James E, Roach, David, Vander Horn, Peter B, Voss, Karl O
Format: Article
Language:English
Subjects:
Analytical chemistry
Chemical and thermal methods
Chemistry
Equipment Design
Exact sciences and technology
Fluorescence
Immunoblotting - instrumentation
Immunoblotting - methods
Isoelectric Focusing
Lasers
Proteins
Spectrometric and optical methods
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Summary:We describe a whole-capillary, multicolor laser-induced fluorescence scanner for microfluidic protein analysis systems. Separation of proteins is achieved by isoelectric focusing in a short length of fused-silica capillary after which the resolved proteins are immobilized to the capillary wall using photochemistry. The capillary is then evacuated, and fluorescently labeled antibodies are flowed through the capillary to bind to the immobilized proteins. This technique provides high sensitivity, the ability to spatially resolve and quantify proteins, and provides the opportunity for complete automation. Results obtained by fluorescence detection are compared to those obtained by chemiluminescence while offering enhanced resolution and signal stability.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac071537z