Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
2001-11-09
2004-01-06
Whisenant, Ethan (Department: 1637)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C435S091200, C536S024300, C536S025320
Reexamination Certificate
active
06673550
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to probes comprising fluorescent compounds. The present compositions may be employed in the detection and quantitation of analytes such as polypeptides, polynucleotides, hormones and drugs. In particular, the invention pertains to fluorescent compounds and their use as detection groups in electrophoretic probes.
BACKGROUND OF THE INVENTION
The need to determine many analytes (for example multiple pathogens or multiple genes or multiple genetic variants) in blood or other biological fluids has become increasingly apparent in many branches of medicine. Most multi-analyte assays, such as multiplexed assays that detect, e.g., multiple nucleic acid sequences, involve multiple steps, have poor sensitivity, a limited dynamic range (typically on the order of 2 to 100-fold differences and some require sophisticated instrumentation.
Fluorescent compounds are widely used as labels in a variety of diagnostic procedures. Fluorescent compounds are linked to other entities to form probes that are used to assay for a wide variety of materials of interest. In some applications it is important that fluorescent compounds employed as labels have different spectral properties. Such properties include wavelength of excitation, wavelength of emission, quantum yields, Stokes shift, and so forth.
BRIEF DESCRIPTION OF THE RELATED ART
U.S. Pat. No. 6,008,379 discloses aromatic substituted xanthene dyes.
U.S. Pat. No. 6,080,852 discusses 4,7-dichlororhodamine dyes.
Unsymmetrical fluorescein derivatives are disclosed in U.S. Pat. No. 4,439,356.
Xanthene dyes having a fused (C) benzo ring are discussed in U.S. Pat. No. 4,945,171.
4,7-Dichlorofluorescein dyes as molecular probes are discussed in WO 94/05688.
U.S. Pat. No. 4,351,760 discloses alkyl substituted fluorescent compounds and polyamino acid conjugates.
U.S. Pat. No. 4,318,846 discusses ether substituted fluorescein polyamino acid compounds as fluorescers and quenchers. PCT WO 97/39064 discloses fluorinated xanthene derivatives.
U.S. Pat. No. 5,807,682 describes probe compositions for detecting a plurality of nucleic acid targets.
SUMMARY OF THE INVENTION
In one embodiment the present invention is directed to electrophoretic tag probes, and sets of such probes, wherein a probe comprises a releasable portion that is separately detectable on the basis of mobility and a detectable moiety that is a compound of the formula:
wherein:
Z is H, lower alkyl, substituted lower alkyl, lower alkenyl, substituted lower alkenyl, lower alkynyl, substituted lower alkynyl, cycloalkyl, alkoxy, substituted alkoxy, phenoxy, substituted phenoxy, aromatic, substituted aromatic, phenyl, substituted phenyl, polycyclic aromatic, substituted polycyclic aromatic, heterocyclic, substituted heterocyclic, chlorine, fluorine, bromine, iodine, COOH, carboxylate, amide, nitrile, nitro, sulfonyl, sulfate, sulfone, amino, tethered amino, quaternary amino, imino, phosphorus containing species such as, e.g., phosphate, phosphite, and the like, polymer chains of from about 2 to about 500, preferably, about 5 to about 100 monomer units such as, e.g., nucleotides, amino acids, amides, esters, ethers, amines, and so forth,
A is O, N
+
(R
1
)(R
2
) wherein R
1
and R
2
are independently H, lower alkyl, substituted lower alkyl, and the like,
D is OH, OR
3
wherein R
3
is lower alkyl, substituted lower alkyl, aryl, substituted aryl, and the like, N(R
1
)(R
2
) wherein R
1
and R
2
are independently H, lower alkyl, substituted lower alkyl,
W
1
, W
2
, W
3
, W
4
, W
5
and W
6
are independently H, lower alkyl, substituted lower alkyl, lower alkenyl, substituted lower alkenyl, lower alkynyl, substituted lower alkynyl, cycloalkyl, alkoxy, substituted alkoxy, phenoxy, substituted phenoxy, aromatic, substituted aromatic, phenyl, substituted phenyl, polycyclic aromatic, substituted polycyclic aromatic, heterocyclic, substituted heterocyclic, chlorine, fluorine, bromine, iodine, COOH, carboxylate, amide, nitrile, nitro, sulfonyl, sulfate, sulfone, amino, tethered amino, quaternary amino, imino, and so forth,
X
1
-X
4
are independently H, lower alkyl, substituted lower alkyl, lower alkenyl, substituted lower alkenyl, lower alkynyl, substituted lower alkynyl, cycloalkyl, alkoxy, substituted alkoxy, phenoxy, substituted phenoxy, aromatic, substituted aromatic, phenyl, substituted phenyl, polycyclic aromatic, substituted polycyclic aromatic, heterocyclic, substituted heterocyclic, chlorine, fluorine, bromine, iodine, COOH, carboxylate, amide, nitrile, nitro, sulfonyl, sulfate, sulfone, amino, tethered amino, quaternary amino, imino, and so forth,
wherein W
2
and W
3
may be taken together to form one or more rings comprising 4 to 14 atoms, preferably, 4 to 8 atoms, more preferably, 5 to 7 atoms, usually carbon atoms, and comprising 1 to 7 unsaturations, usually, 1 to 4 unsaturations, such as, e.g., benzo (from benzene), naptho (from naphthalene), anthro (from anthracene), and the like, and
wherein W
4
and W
5
may be taken together to form a ring comprising 4 to 14 atoms, preferably, 4 to 8 atoms, more preferably, 5 to 7 atoms, usually carbon atoms, and comprising 1 to 7 unsaturations, usually, 1 to 4 unsaturations, such as, e.g., benzo (from benzene), naptho (from naphthalene), anthro (from anthracene), and the like,
The electrophoretic probes may be used in methods for simultaneously determining multiple analytes in a sample suspected of containing the analytes. In the method a combination is provided comprising the sample and an electrophoretic tag probe comprising a releasable portion that is separately detectable on the basis of mobility and the presence therein of a fluorescent compound in accordance with the invention. The electrophoretic tag probe, or e-tag probe, is involved in a binding event that is related to the presence of the analyte in the sample. The combination is treated with reagents under conditions sufficient to release the releasable portion, forming an e-tag reporter. The presence and/or amount of the released e-tag reporter is detected and is related to the presence and/or amount of the analyte in the sample.
REFERENCES:
patent: 4274240 (1981-06-01), Soum
patent: 4331590 (1982-05-01), Bocuslaski et al.
patent: 4650750 (1987-03-01), Giese
patent: 4675300 (1987-06-01), Zare et al.
patent: 4709016 (1987-11-01), Giese
patent: 4780421 (1988-10-01), Kameda
patent: 5324401 (1994-06-01), Yeung et al.
patent: 5340716 (1994-08-01), Ullman
patent: 5360819 (1994-11-01), Giese
patent: 5470705 (1995-11-01), Grossman et al.
patent: 5470967 (1995-11-01), Huie et al.
patent: 5516636 (1996-05-01), McCapra
patent: 5516931 (1996-05-01), Giese
patent: 5536834 (1996-07-01), Singh et al.
patent: 5552028 (1996-09-01), Madabhushi et al.
patent: 5560811 (1996-10-01), Briggs et al.
patent: 5565324 (1996-10-01), Still et al.
patent: 5573906 (1996-11-01), Bannwarth et al.
patent: 5580732 (1996-12-01), Grossman et al.
patent: 5604104 (1997-02-01), Giese
patent: 5610020 (1997-03-01), Giese
patent: 5622929 (1997-04-01), Willner
patent: 5624800 (1997-04-01), Grossman et al.
patent: 5650270 (1997-07-01), Giese
patent: 5703222 (1997-12-01), Grossman et al.
patent: 5709994 (1998-01-01), Pease
patent: 5719028 (1998-02-01), Dahlberg et al.
patent: 5721099 (1998-02-01), Still et al.
patent: 5723591 (1998-03-01), Livak et al.
patent: 5756726 (1998-05-01), Hemmi et al.
patent: 5789172 (1998-08-01), Still et al.
patent: 5807675 (1998-09-01), Davalian
patent: 5811239 (1998-09-01), Frayne
patent: 5843655 (1998-12-01), McGall
patent: 5843666 (1998-12-01), Akhavan-Tafti et al.
patent: 5846839 (1998-12-01), Gallop
patent: 5874213 (1999-02-01), Cummins et al.
patent: 5876930 (1999-03-01), Livak et al.
patent: 5989871 (1999-11-01), Grossman et al.
patent: 5998140 (1999-12-01), Dervan et al.
patent: 6001579 (1999-12-01), Still et al.
patent: 6027890 (2000-02-01), Ness
patent: 6045676 (2000-04-01), Mathies et al.
patent: 6090947 (2000-07-01), Dervan et al.
patent: 6251581 (2001-06-01), Ullman et al.
patent: 6312893 (2001-11-01), Van Ness
patent: 6368874 (2002-04-
Hernandez Vincent
Matray Tracy
Singh Sharat
Aclara Biosciences, Inc.
Macevicz Stephen C.
Tung Joyce
Whisenant Ethan
LandOfFree
Electrophoretic tag reagents comprising fluorescent compounds does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Electrophoretic tag reagents comprising fluorescent compounds, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Electrophoretic tag reagents comprising fluorescent compounds will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3187050