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Flow Cytometry-Based Biosensor for Detection of Multivalent Proteins

✍ Scribed by Xuedong Song; Jeane Shi; Basil Swanson

Publisher
Elsevier Science
Year
2000
Tongue
English
Weight
90 KB
Volume
284
Category
Article
ISSN
0003-2697

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✦ Synopsis

Microsphere

-based flow cytometric detection of cholera toxin (CT) through distance-dependent fluorescence resonant energy transfer (FRET) has been developed. Simultaneous double-fluorescence changes induced by multivalent interactions between CT and fluorophore (both fluorescence donor and acceptor)labeled ganglioside GM1 on a biomimetic membrane surface (supported bilayers of phospholipids) can be measured by a commercial flow cytometer, providing a convenient and sensitive detection method for CT. The flow cytometry-based biosensor is capable of detecting less than 10 pM CT within 30 min. The signal generation strategy coupled with flow cytometry also provides a convenient method for kinetic studies of multivalent interactions. The surface density and the ratio of donor/acceptor-labeled GM1 on the surfaces of phospholipid bilayers are optimized to achieve high sensitivity.

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