Beckman Coulter, Inc.

Beckman Instruments, Inc.

Other

M Parameswara Reddy

Firdous Farooqui

Daniel A Keys

Naeem B Hanna

Chitra Kumari Ratnayake

Maged A Michael

Yu Liu

Jirong Gu

Tung-Liang Huang

James C Sternberg

Kurt L Brillhart

Robert Bruce Wallace

Edward V Koh

Tung-Lian Huang

Adrian Clausell

Methods for determining the presence of an analyte in a sample are disclosed, in which a capture agent is bound to the analyte at a first epitope and a detection agent is bound at a second epitope, and in which the detection agent includes an oligonucleotide to which a labeled, complementary oligonucleotide can be hybridized.

Labeled complementary oligonucleotides to detect oligonucleotide-linked ligands

This invention relates to improved methods and compositions for conducting Capillary Electrophoresis (CE) to separate molecules on the basis of their respective size or charge.

Methods and compositions for capillary electrophoresis (CE)

This invention relates to spotting solutions for the production of assay articles with uniform spot size and morphology for the detection of biopolymers, comprising N-lauroyl sarcosine in a buffered solution. Optionally the spotting solution may further comprise a fluorophore or a dye. This invention further provides means for non-destructive quality control of the production of said assay articles. This invention further provides means for non-destructive quality control of the production of said assay articles.

Formulation of spotting solution to achieve uniform spot size and morphology and for nondestructive quality control of assay articles

The present invention provides a useful system for assays that comprises a solid support, a plurality of capture oligonucleotides immobilized onto the solid support, and complementary oligonucleotides attached to capture ligands. A detectable label can be directly attached to the capture oligonucleotides or the complementary oligonucleotides. The labeled oligonucleotides can be detected, and used to determine the quality of the assay. A labeled detector ligand corresponding to a target ligand can also be independently detected apart from the labeled oligonucleotide.

Assay system using labeled oligonucleotides

The ability to efficiently determine the state of enzyme expression in cells has long been desired as material to the diagnosis of disease. This invention relates to cytoenzymology, and more particularly to improved reagents for use in cell-based assays, especially those using fluorogenic substrates.

Methods and reagents for improved cell-based assays

Oligonucleotide probes, kits, and methods useful for detecting a polynucleotide target in a sample are provided. The method, a mixture is formed by combining a polynucleotide target sample, a first probe that is complementary to the polynucleotide target and having a first fluorescent donor or fluorescent acceptor; and a second probe that is partially complementary to the first probe and having a second fluorescent donor or fluorescent acceptor. The second probe competes with the polynucleotide target for binding to the first probe and the first probe preferentially binds to the polynucleotide target rather than to the second probe. The-first fluorescent donor or acceptor and second fluorescent donor or acceptor form a donor/acceptor pair capable of fluorescence resonance energy transfer (FRET) with each other in response to activation of the fluorescent donor by light of a predetermined wavelength or band of wavelengths.

Oligonucleotide probes for detecting nucleic acids through changes in flourescence resonance energy transfer

Formamide with low conductivity and a neutral or slightly basic pH is provided. Such formamide may be used in a sample loading solution for capillary electrophoretic separation of biomolecular analytes to enhance the efficiency of sample injection into the capillary and the intensity of the signal. Formamide of the present invention may be obtained by first purifying the formamide to a conductivity of below 7 micro mho, and then adjusting the pH of the purified formamide with a base to a range of about pH 6.5 to 7.5. The purification may be carried out by first removing the water from the formamide, and then distilling the dried formamide until the conductivity of formamide is below about 7 micro mho.

Reagents for preparing a sample containing biomolecular analytes for capillary electrophoretic separation and methods for making the reagents

An article of manufacture is provided that is useful in differentiating between solutes, such as during electrophoretic separations. An embodiment of the article is a capillary tube, that carries a polymer along the inner wall of the capillary tube. The polymer is effective to reduce undesired interactions and preferably includes a polylactam that is absorbed to the surface before the surface is exposed to the solutes. A preferred polylactam is poly(vinylpyrrolidone) with a molecular weight greater than about 1,000,000 daltons (weight-average).

Surfaces with reduced electroosmotic flow

Efficient cyclic-bridged cyanine dyes

Reagents and methods for preparing samples containing both biomolecule analytes and macrobiomolecules for capillary electrophoresis separation are provided. The reagents comprise a branched polymer. When mixed with a sample containing both biomolecule analytes and macrobiomolecules, particularly DNA templates, the branched polymer of the reagent can suppress the entrance of the macrobiomolecules into a capillary electrophoresis tube during electrokinetic injection of the sample into the capillary electrophoresis tube.

Method and reagent for suppressing the injection of nucleic acid sequencing template into micro-bore capillary during DNA separation with capillary electrophoresis

Activating groups based on N-hydroxynaphthalimide, are disclosed herein. The activating groups can mediate the coupling of labeling moieties, such as biotin or cyanine dyes, to a variety of components, including chain terminators, nucleoside triphosphates, and oligonucleotides, which are used in nucleotide sequencing. From these activating groups, activated esters of the labeling moieties can be prepared. The activated esters react with a component, for example a derivatized nucleotide chain terminator, to give a labeled component. In additions, methods of the present invention provide for labeling a nucleoside triphosphate in organic media. The activating groups and methods of the present invention allow the activation and coupling reactions to occur at a much higher yield, compared with the prior art.

Efficient activated cyanine dyes

Universal solid support oligonucleotide synthesis reagents, oligonucleotide synthesis processes, and reagents for cleaving oligonucleotides from solid supports are disclosed. Oligonucleotide synthesis reagents have the following general formula:

Universal solid supports and methods for their use

Disclosed herein are N.sup.4 protected deoxycytidines for use in the synthesis of oligonucleotides, the protecting groups being represented by the formula: --CO--(CH.sub.2).sub.0-9 --CH.sub.3. Preferred embodiments are N.sup.4 acetyl deoxycytidines and include N.sup.4 acetyl deoxycytidine phosphoramidites and N.sup.4 acetyl deoxycytidine H-phosphonates. When used to prepare oligonucleotides the protected deoxycytidine compounds provide high quality oligonucleotide products with little side product from cleavage and deprotection reactions carried out with alkyl amine compounds.

CAC H-phosphonate and its use in the synthesis of oligonucleotides

Double stranded nucleic acid duplexes serve as universal harvestable and cleavable link systems in a variety of different types of immunoassays (e.g., sandwich, competitive, etc.). Depending upon the type of assay, at least one specific component involved in the assay system is attached to a first member of a pair of sequences forming a double stranded nucleic acid (i.e., two oligonucleotides comprising substantially complementary sequences). The assay is carried out in the presence of a support to which is attached an oligonucleotide which is the other member of the pair of sequences forming a double-stranded nucleic acid duplex under hybridization conditions. Upon the hybridization of the two complementary oligonucleotides to form a duplex, the component of the assay system to which the first member of the pair of oligonucleotides is attached may thereby be effectively removed from the solution phase and harvested onto the support. Oligonucleotides bound to a support are reusable in multiple successive assays. Moreover, any given support-bound oligonucleotide can be used in accordance with the present invention for the analysis of a variety of different analytes. In many cases, the assay system includes a label to facilitate quantifying the amount of analyte; in others, the amount of analyte may be determined without the use of any extraneous label.

Compositions and methods for use in detection of analytes

Deoxyribonucleotide and ribonucleotide derivatives of the general formula I ##STR1## wherein R.sup.1 represents --CR'R'--Ar, in which Ar is substituted aryl (as hereinafter defined) and R' and R' are independently selected from the group consisting of hydrogen and lower alkyl; one of --R.sup.2 and R.sup.3 is a hydroxyl-protecting group and the other is a group suitable for synthesis of polynucleotides or for attachment of the nucleotide to a solid support; R.sup.4 is selected from the group consisting of hydrogen, --OH and protected hydroxyl; and B represents a divalent radical corresponding to a purine or pyrimidine base. When synthesis is carried out using these derivatives, the deprotection procedure is reduced to an essentially instantaneous process. The derivatives have acceptable shelf life and are very stable to conventional DNA synthesis conditions. Particularly preferred are those compounds wherein Ar is mono- and dihalo-substituted phenyl.

Growing community of inventors

Brea, CA, United States of America

M Parameswara Reddy