Resonant energy transfer based biosensor for detection of multivalent proteins.

Conference · Mon Jan 01 00:00:00 EST 2001

DOI:https://doi.org/10.1117/12.372905· OSTI ID:975774

Song, X ^[1]; Swanson, Basil I

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1. Xuedong

We have developed a new fluorescence-based biosensor for sensitive detection of species involved in a multivslent interaction. The biosensor system utilizes specific interactions between proteins and cell surface receptors, which trigger a receptor aggregation process. Distance-dependent fluorescence self-quenching and resonant energy transfer mechanisms were coupled with a multivalent interaction to probe the receptor aggregation process, providing a sensitive and specific signal transduction method for such a binding event. The fluorescence change induced by the aggregation process can be monitored by different instrument platforms, e.g. fluorimetry and flow cytometry. In this article, a sensitive detection of pentavalent cholera toxin which recognizes ganglioside GM1 has been demonstrated through the resonant energy transfer scheme, which can achieve a double color change simultaneously. A detection sensitivity as high as 10 pM has been achieved within a few minutes (c.a. 5 minutes). The simultaneous double color change (an increase of acceptor fluorescence and a decrease of donor fluorescence intensity) of two similar fluorescent probes provides particularly high detection reliability owing to the fact that they act as each other's internal reference. Any external perturbation such as environmental temperature change causes no significant change in signal generation. Besides the application for biological sensing, the method also provides a useful tool for investigation of kinetics and thermodynamics of a multivalent interaction. Keywords: Biosensor, Fluorescence resonant energy transfer, Multivalent interaction, Cholera Toxin, Ganglioside GM1, Signal Transduction

• Research Organization: Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)
• Sponsoring Organization: USDOE
• OSTI ID: 975774
• Report Number(s): LA-UR-00-5531; TRN: US201018%%861
• Resource Relation: Journal Volume: 3858; Conference: Submitted to: SPIE Conference on Advanced Materials and Optical Systems for Chemical 2 and Biological Detection, Boston, Massachusetts, September 1999
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE
AMBIENT TEMPERATURE
CHOLERA
COLOR
DETECTION
ENERGY TRANSFER
FLUORESCENCE
FLUORESCENCE SPECTROSCOPY
GANGLIOSIDES
KINETICS
OPTICAL SYSTEMS
PROBES
PROTEINS
RELIABILITY
SENSITIVITY
THERMODYNAMICS
TOXINS