Enzo Diagnostics, Inc.

Enzo Life Sciences, Inc. C/o Enzo Biochem, Inc.

Enzo Biochem, Inc.

Stanley Kline

Elazar S Rabbani

Dean L Engelhardt

Dollie Kirtikar

Jannis G Stavrianopoulos

Janes G Stavrianopoulos

A process for determining the sequence of nucleic acids of interest employs nucleotides or nucleotide analogs that have been made detectable by non-radioactive modifying or labeling. Such nucleotides or nucleotide analogs are modified on the sugar moieties, the phosphate moieties or the base moieties, including base analogs. Modified nucleotide analogs can be attached to or coupled to or incorporated into DNA or RNA. The modified or labeled nucleotides or nucleotide analogs are also useful in processes for detecting the presence of nucleic acids of interest and for characterizing chromosomal sequences. Detection processes using the modified or labeled nucleotides or nucleotide analogs extend to the use of a gel for separating or resolving hybrids formed between non-radioactively labeled oligonucleotides or polynucleotides and such nucleic acids of interest.

Nucleic acid sequencing processes using non-radioactive detectable modified or labeled nucleotides or nucleotide analogs, and other processes for nucleic acid detection and chromosomal characterization using such non-radioactive detectable modified or labeled nucleotides or nucleotide analogs

The present invention provides a nucleotide having the formula, wherein PM is a phosphate moiety, SM is a ribose or a deoxyribose sugar moiety, and BASE is a pyrimidine, purine or 7-deazapurine moiety. PM is attached to SM at a position independently selected from the 2', 3&#x2032;, and 5&#x2032; positions of SM when the nucleotide is a ribonucleotide, and at a position independently selected from the 3&#x2032; and 5&#x2032; positions when the nucleotide is a deoxyribonucleotide. BASE is attached to the 1&#x2032; position of SM from the Nposition when BASE is a pyrimidine, or the Nposition when BASE is a purine or 7-deazapurine. Sig is a detectable moiety covalently attached to SM directly or through a linkage group. Also provided are an oligo- or polynucleotide comprising at least one such sugar moiety labeled nucleotide, and other compositions including those wherein a polypeptide is terminal ligated or attached to the oligo- or polynucleotide. The sugar moiety labeled nucleotide, and the oligo- or polynucleotides and other compositions comprising such nucleotides, are useful as diagnostic tools for detecting analytes and as therapeutic agents.

Sugar moiety labeled nucleotide, and an oligo- or polynucleotide, and other compositions comprising such sugar moiety labeled nucleotides

The present invention provides a nucleotide having the formula,Sig-PM-SM-BASEwherein PM is a phosphate moiety, SM is a sugar moiety and BASE is a pyrimidine, purine or 7-deazapurine moiety. PM is attached to the 3' or the 5&#x2032; position of the sugar moiety when the nucleotide is a deoxyribonucleotide and at the 2&#x2032;, 3&#x2032; or 5&#x2032; position when the nucleotide is a ribonucleotide. BASE is attached to the 1&#x2032; position of SM from the Nposition when BASE is a pyrmidine or the Nposition when BASE is a purine or 7-deazapurine. Sig is covalently attached to PM directly or via a chemical linkage, and represents a detectable moiety covalently attached to SM directly or through a linkage group. Also provided are an oligo- or polynucleotide comprising at least one such phosphate-moiety labeled nucleotide, and other compositions including those wherein a polypeptide is terminally ligated or attached to the oligo- or polynucleotide. The phosphate-moiety labeled nucleotide, and the oligo- or polynucleotides and other compositions comprising such phosphate-moiety labeled nucleotides, are useful as diagnostic tools for detecting analytes and as therapeutic agents.

Oligo- or polynucleotides comprising phosphate-moiety labeled nucleotides

Disclosed is an assay system including a compound comprising an analyte-specific moiety having substituted thereon a polymer comprising plurality of self-quenching emitter moieties and a plurality of isocharged functionality separating the emitter moieties. The present invention provides compounds that overcome the undesirable effects of self-quenching when multiple emitter moieties are used for labelling of assay reagents. Avoidance of this self-quenching phenomenon by the compounds of the invention makes it possible to introduce a more concentrated degree of labelling on to analyte-specific molecules such as oligo nucleotide probes, antibodies and other specific binding proteins and analyte-specific polysaccharides. Therefor, it is possible to effect greater assay sensitivity because the number of labels per recognition molecule(analyte-specific moiety) can be increased beyond the point previously possible without the reduction in signal caused by self-quenching.

Specific binding assay compound with inhibitive self-quenching

The present invention provides nucleotides and polynucleotides which are chemically modified or labeled so as to be capable of ready detection when attached to and/or incorporated in nucleic acid material. More particularly, this invention provides a nucleotide selected from the group consisting of (i) a ribonucleotide having the formula, ##STR1## wherein PM is a phosphate moiety, SM is a sugar moiety, BASE is a pyrimidine, purine or 7-deazapurine moiety, and Sig is a saccharide moiety. PM is attached at the 2',3' or 5' position of SM, BASE is attached to the 1' position of SM from the N.sup.1 position when BASE is a pyrimidine or covalently attached to SM; and (ii) a nucleotide having the formula, ##STR2## wherein PM, SM, BASE and Sig are as defined above but PM is attached to the 3' or the 5' position of SM when said nucleotide is a deoxyribonucleotide and at the 2', 3' or 5' position when said nucleotide is a ribonucleotide, BASE is attached to the 1' position of SM from the N.sup.1 position when BASE is a pyrimidine or the N.sup.9 position when BASE is a purine, and wherein Sig is covalently attached to PM.

Saccharide specific binding system labeled nucleotides

The present invention provides nucleotides and polynucleotides which are chemically modified or labeled so as to be capable of ready detection when attached to and/or incorporated in nucleic acid material. More particularly, this invention provides a nucleotide having the formula

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Brooklyn, NY, United States of America

Stanley Kline