Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Reexamination Certificate
1999-10-27
2003-04-01
Whisenant, Ethan (Department: 1655)
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
C536S063000, C536S063000, C435S006120, C436S094000
Reexamination Certificate
active
06541617
ABSTRACT:
FIELD OF THE INVENTION
The invention relates to the use of particles comprising binding ligands and electron transfer moieties (ETMs). Upon binding of a target analyte, a particle and a reporter composition are associated and transported to an electrode surface. The ETMs are then detected, allowing the presence or absence of the target analyte to be determined.
BACKGROUND OF THE INVENTION
There are a number of assays and sensors for the detection of the presence and/or concentration of specific substances in fluids and gases. Many of these rely on specific ligand/antiligand reactions as the mechanism of detection. That is, pairs of substances (i.e. the binding pairs or ligand/antiligands) are known to bind to each other, while binding little or not at all to other substances. This has been the focus of a number of techniques that utilize these binding pairs for the detection of the complexes. These generally are done by labeling one component of the complex in some way, so as to make the entire complex detectable, using, for example, radioisotopes, fluorescent and other optically active molecules, enzymes, etc.
Other assays rely on electronic signals for detection. Of particular interest are biosensors. At least two types of biosensors are known; enzyme-based or metabolic biosensors and binding or bioaffinity sensors. See for example U.S. Pat. No. 4,713,347; 5,192,507; 4,920,047; 3,873,267; and references disclosed therein. While some of these known sensors use alternating current (AC) techniques, these techniques are generally limited to the detection of differences in bulk (or dielectric) impedance, and rely on the use of mediators in solution to shuttle the charge to the electrode.
Recently, there have been several preliminary reports on the use of very short connections between a binding ligand and the electrode, for direct detection, i.e. without the use of mediators. See Lbtzbeyer et al., Bioelectrochemistry and Bioenergetics 42:1-6 (1997); Dong et al., Bioelectrochemistry and Bioenergetics 42:7-13 (1997).
In addition, there are a number of reports of self-assembled monolayers of conjugated oligomers on surfaces such as gold. See for example Cygan et al., J. Am. Chem. Soc. 120:2721 (1998).
In addition, Charych et al. report on the direct colormetric detection of a receptor-ligand interaction using
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bilayer assembly (Science 261:585 (1993).
PCT applications WO 95/15971, PCT/US96/09769, PCT/US97/09739, W096/40712 and PCT/US97/20014 describe novel compositions comprising nucleic acids containing electron transfer moieties, including electrodes, which allow for novel detection methods of nucleic acid hybridization.
In addition, there are a number of sensors that rely on the use of particles, including magnetic particles, particularly for electrochemiluminescence detection. See U.S. Pat. Nos. 5,746,974; 5,770,459; 5,779,976; 4,731,337; 4,115,535; 4,777,145; 4,945,045; 4,978,610; 5,705,402; 4,910,148; 5,512,439; 5,585,241; and 5,609,907; and WO 90/14891; WO 90/05301; WO 92/14139; and WO 90/06044.
Finally, there are reports that bring particles together using nucleic acids for archetectural reasons. See Mirkin et al., Nature 382:607 (1996); Mirkin et al., Science 277:1078 (1997); Storhoff et al., J. Am. Chem. Soc. 120:1959 (1998); and WO 98/10289.
SUMMARY OF THE INVENTION
In accordance with the objects outlined above, the present invention provides compositions comprising a gold colloid particle comprising at least one ETM. The colloid can further comprise a self-assembled monolayer (SAM). The compositions can further comprise an electrode, that may also contain a SAM. In an additional aspect, the invention provides compositions comprising gold colloid particles comprising a SAM, a capture probe, an amplification sequence; and a label probe hybridized to the amplification sequence, wherein the label probe comprises at least one covalently attached ETM.
In an additional aspect, the invention provides compositions comprising a transport composition comprising a first binding partner that directly or indirectly binds a target analyte, and a reporter composition. The reporter composition comprises a second binding partner that directly or indirectly binds the target analyte and a plurality of electron transfer moieties (ETMs). At least one of the transport and the reporter compositions is a particle. Upon introduction of the target analyte, the transport composition and the reporter composition are associated. These compositions may also comprise SAMs, and an electrode, optionally containing a SAM, can be included.
In a further aspect, the invention provides methods of detecting a target analyte in a sample. The methods comprise adding the sample to a composition as outlined above, such that the target analyte binds to the transport composition and the reporter composition to form an assay complex. The assy
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complex is transported to the electrode, and the presence or absence of the ETMs is detected.
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Bamdad Cynthia C.
Mucic Robert C.
Clinical Micro Sensors, Inc.
Flehr Hohbach Test Albritton & Herbert LLP
Lu Frank
Silva Robin M.
Trecartin Richard F.
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