Prolinx Incorporated

Agilent Technologies, Inc.

California Institute of Technology

Systemix, Inc.

Other

Cambrex Bio Science Rockland, Inc.

Porlinx, Incorporated

Mark L Stolowitz

Robert J Kaiser

Kevin P Lund

Karin A Hughes

Edward A Kesicki

Jean P Wiley

Guisheng Li

Clarence N Ahlem

Steven M Torkelson

Douglas A Spicer

James E Mahoney

Amy L Springer

Carl H D Weissman

Sidney F Cook

The present invention provides a sensor surface comprising: a substrate coated with a free electron metal; and a matrix layer disposed on the free electron metal, wherein the matrix layer comprises an organic compound having a boronic acid complexing moiety. The matrix is preferably a self-assembled monolayer (SAM), a mixed self-assembled monolayer (mSAM), or combinations thereof.

Sensor surfaces for detecting analytes

The present invention provides a method for detecting an analyte, comprising providing a sensor comprising a substrate coated with a free electron metal, a matrix layer disposed on the free electron metal, the matrix layer comprising an organic compound, wherein the organic compound has a boronic acid complexing moiety and, a boronic acid moiety complexed to the boronic acid complexing moiety; and contacting the sensor with the analyte to elicit a response; and measuring the response thereby detecting the analyte.

Sensor surfaces for detecting analytes and methods of use

A reagent having the general formula of General Formula I:wherein group R is an electrophilic or a nucleophilic moiety suitable for reaction of the reagent with a biologically active species; group Ris one of H and OH moieties; group Ris one of an alkyl and a methylene bearing an electronegative substituent; group Z is one of (CH)and CHO(CHCHO), and n is an integer of from 1 to 5, and nis an integer of from 1 to 4; and each of group Zand Zare (CH)CONH(CH)CONH(CH). Also, a reagent having the general formula of General Formula II:group Ris one of H and OH moieties; group Z is one of (CH)and CHO(CHCHO), and n is an integer of from 1 to 5, and nis an integer of from 1 to 4; each of group Zand group Zinclude one of (CH)CONH(CH)CONH(CH); and (CH)CONH(CH); and group R is an electrophilic or a nucleophilic moiety suitable for reaction of the reagent with the biologically active species.

Bifunctional boronic compound complexing reagents and complexes

This invention provides methods for removing unincorporated dye-labeled molecules from a mixture that includes dye-labeled polynucleotides or other polymers and the unincorporated dye-labeled molecules. The methods involve adsorbing the unincorporated dye-labeled molecules into a plurality of particles that are made up of one or more porous hydrophobic materials that are encapsulated in a hydrophilic matrix.

Removal of dye-labeled dideoxy terminators from DNA sequencing reactions

1,2-Phenylenediboronic acid reagents and complexes

Phenyldiboronic acid reagents and complexes

This invention provides methods for removing unincorporated fluorescent dye-labeled molecules from a mixture that includes fluorescently-labeled polynucleotides and the unincorporated fluorescent dye-labeled molecules. The methods involve adsorbing the unincorporated fluorescent dye-labeled molecules into a plurality of particles that are made up of one or more porous hydrophobic materials that are encapsulated in a hydrophilic matrix.

Reagents suitable for the modification of a bioactive species for the purpose of incorporating a bifunctional boronic compound complexing moiety for subsequent conjugation to a different (or the same) bioactive species having pendant phenylboronic acid moieties of General Formula 1, ##STR1## wherein group R is an electrophilic or nucleophilic moiety suitable for reaction of the putative bifunctional boronic compound complexing reagent with a bioactive species, wherein group R.sub.2 is selected from one of H and OH moieties, and wherein group R.sub.3 is selected from one of an alkyl and a methylene bearing an electronegative substituent. Group Z is a spacer selected from (CH.sub.2).sub.n and CH.sub.2 O(CH.sub.2 CH.sub.2 O).sub.n.sbsb.2, wherein n is an integer of from 1 to 5, and wherein n.sub.2 is an integer of from 1 to 4. Each of group Z.sub.2 and Z.sub.3 is a spacer selected from CH.sub.2 Ar, CH.sub.2 CONHCH.sub.2 Ar, CH.sub.2 CONH(CH.sub.2).sub.n.sbsb.3 CO--NHCH.sub.2 Ar, and (CH.sub.2).sub.n.sbsb.4 NHCO(CH.sub.2).sub.n.sbsb.5 CONHCH.sub.2 Ar, wherein the group Ar represents the aromatic ring in the reagent of General Formula I to which the spacer Z.sub.2 or Z.sub.3 is appended, wherein n.sub.3 is an integer of from 1 to 5, wherein n.sub.4 is an integer selected from one of 2 and 3, and wherein n.sub.5 is an integer of from 1 to 4. Reaction of a reagent of General Formula I with a bioactive species affords a conjugate having pendant putative bifunctional boronic compound complexing moieties. (one or more) The conjugate may be further reacted with hydroxylamine (NH.sub.2 OH) by amidation of the benzoic acid ester moiety to afford a class of bifunctional boronic compound complexing conjugate, e.g., conjugate with one or more pendant bifunctional boronic compound complexing moieties.

Reagents suitable for the modification of a bioactive species for the purpose of incorporating one or more phenyldiboronic acid (PDBA) moieties for subsequent conjugation to a different (or the same) bioactive species having one or more pendant boronic compound complexing moieties of the general formula of General Formula I, ##STR1## wherein group R is a reactive electrophilic or nucleophilic moiety suitable for reaction of the PDBA reagent with a bioactive species. Group Z is a spacer selected from a saturated or unsaturated chain up to about 0 to 6 carbon equivalents in length, an unbranched saturated or unsaturated chain of from about 6 to 18 carbon equivalents in length with at least one intermediate amide or disulfide moiety, and a polyethylene glycol chain of from about 3 to 12 carbon equivalents in length. Group Q is a linkage that includes one of amide, ether and thioether moieties. Group R is preferably selected from, but not limited to, one of acrylamide, bromo, bromoacetamide, chloro, chloroacetamide, dithiopyridyl, hydrazide, N-hydroxysuccinimidyl ester, N-hydroxysulfo-succinimidyl ester, imidate ester, imidazolide, iodo, iodoacetamide, maleimide, amino and thiol moieties. Group Z is preferably an unbranched alkyl chain of the general formula (CH.sub.2)n, wherein n=1 to 6. Group Q is preferably selected from one of NHCO, CONH, NHCOCH.sub.2, CONHCH.sub.2, O, OCH.sub.2, S and SCH.sub.2 moieties.

Boron compound complexing reagents, boron compound complexes, and methods of synthesizing these reagents and complexes are disclosed. These reagents and complexes include those shown in General Formula CIII, General Formula CIV, and General Formula CVI. In one embodiment, the reagents of General Formula CIII may be used to produce, after condensation with a bioactive species (BAS), the reagent of General Formula CIV. The reagent of General Formula CIV may be used to form a complex with a boron compound, such as a complex shown in General Formula CVI. ##STR1##

The present invention provides arylboronic acid reagents useful for incorporation into modified oligonucleotides and polynucleotides. The modified oligonucleotides and polynucleotides thus produced are useful in bioconjugation reactions for the immobilization and purification of macromolecules.

Boronic acid containing phosphoramidite reagents

The present invention relates to the field of nucleic acid immobilization, purification and detection and, more particularly, to boronic acid modified oligonucleotides and polynucleotides useful in bioconjugation reactions. The modified oligonucleotides and polynucleotides are useful in reactions for the immobilization and purification of macromolecules.

Boronic acid containing oligonucleotides and polynucleotides

The present invention relates to novel class of phenylboronic acid complexing reagents useful for the preparation of bioconjugates, and the method of making and using such reagents. In the present invention, in the place of prior art Avidin-Biotin and Digoxigenin-anti-Digoxigenin systems, phenylboronic acid complexing reagents are utilized in conjunction with phenylboronic acid reagents (many of which are known in the prior art) to facilitate chemical conjugation without the use of intermediary biological macromolecules. Reagents suitable for the modification of a bioactive species for the purpose of incorporating a phenylboronic acid complexing moiety for subsequent conjugation to a different bioactive species having pendant phenylboronic acid moieties are of General Formula I or General Formula II. ##STR1## wherein group X is selected from either H, OH, NH.sub.2, NHCH.sub.3, NHOH and NHOCH.sub.3, wherein group Y is selected from either O, S and NH, and wherein group Z comprises a spacer which either separates the boronic acid complexing moiety from group R, as in General Formula I, or seperates two boronic acid complexing moieties, as in General Formula II. Group R is a reactive electrophilic or nucleophilic moiety suitable for reaction of the phenylboronic acid complexing reagent with a bioactive species.

Phenylboronic acid complexing reagents derived from aminosalicylic acid

Modified nucleotides and polynucleotides which are useful in hybridization assays for the detection of target genes are provided. The modified polynucleotides contain at least one boronic acid moiety which is attached to a nucleotide base in a position which does not interfere with the hydrogen bonding capabilities of that base during duplex formation. The modified polynucleotides are typically formed from naturally occurring nucleotides and one or more modified nucleotides.

Growing community of inventors

Woodinville, WA, United States of America

Mark L Stolowitz