公开(公告)号：US07396647B2

公开(公告)日：2008-07-08

申请号：US11237467

申请日：2005-09-27

Applicant: Elazar Rabbani ,  Jannis G. Stavrianopoulos ,  James J. Donegan ,  Jack Coleman ,  Dakai Liu

Inventor： Elazar Rabbani ,  Jannis G. Stavrianopoulos ,  James J. Donegan ,  Jack Coleman ,  Dakai Liu

IPC: C12Q1/68 ,  C07H21/04

CPC classification number: C12Q1/6818 ,  C12Q1/6834 ,  C12Q1/6844 ,  C12Q1/6853 ,  C12Q1/686 ,  G01N21/64 ,  G01N21/6486 ,  C12Q2565/101 ,  C12Q2533/101 ,  C12Q2561/113

Abstract: This invention provides for compositions for use in real time nucleic acid detection processes. Such real time nucleic acid detection processes are carried out with energy transfer elements attached to nucleic acid primers, nucleotides, nucleic acid probes or nucleic acid binding agents. Real time nucleic acid detection allows for the qualitative or quantitative detection or determination of single-stranded or double-stranded nucleic acids of interest in a sample. Other processes are provided by this invention including processes for removing a portion of a homopolymeric sequence, e.g., poly A sequence or tail, from an analyte or library of analytes. Compositions useful in carrying out such removal processes are also described and provided.

Abstract translation: 本发明提供用于实时核酸检测过程的组合物。 这种实时核酸检测方法是用连接于核酸引物，核苷酸，核酸探针或核酸结合剂的能量转移元件进行的。 实时核酸检测允许定性或定量检测或测定样品中感兴趣的单链或双链核酸。 本发明提供了其它方法，包括从被分析物或分析物库中除去一部分均聚物序列，例如聚A序列或尾部的方法。 还描述和提供了用于进行这种去除方法的组合物。

公开(公告)号：US07323571B2

公开(公告)日：2008-01-29

申请号：US10764389

申请日：2004-01-23

Applicant: Jannis G. Stavrianopoulos ,  Elazar Rabban

Inventor： Jannis G. Stavrianopoulos ,  Elazar Rabban

IPC: C07D293/00 ,  C07D291/00 ,  C07D207/00 ,  C07H21/02 ,  C07H21/04

CPC classification number: C07H21/00 ,  C09B23/02 ,  C09B47/00 ,  C09B57/00 ,  Y10S436/80

Abstract: This invention provides for labeling reagents, labeled targets and processes for preparing labeling reagents. The labeling reagents can take the form of cyanine dyes, xanthene dyes, porphyrin dyes, coumarin dyes or composite dyes. These labeling reagents are useful for labeling probes or targets, including nucleic acids and proteins. These reagents can be usefully applied to protein and nucleic acid probe based assays. They are also applicable to real-time detection processes.

Abstract translation: 本发明提供标记试剂，标记的靶标和制备标记试剂的方法。 标记试剂可以采用花青染料，呫吨染料，卟啉染料，香豆素染料或复合染料的形式。 这些标记试剂可用于标记探针或靶标，包括核酸和蛋白质。 这些试剂可用于基于蛋白质和基于核酸探针的测定。 它们也适用于实时检测过程。

公开(公告)号：US5989809A

公开(公告)日：1999-11-23

申请号：US378118

申请日：1995-01-24

Applicant: Jannis G. Stavrianopoulos

Inventor： Jannis G. Stavrianopoulos

IPC: C07H21/00 ,  C12Q1/68

CPC classification number: C07H21/00 ,  C12Q1/6816

Abstract: The present invention provides a method for detecting the presence of a target polynucleotide in a sample. The method comprises the steps of (a) contacting the sample under hybridizing conditions with (i) a single-stranded polynucleotide probe capable of hybridizing to the target polynucleotide and comprising a polynucleotide and at least one intercalating molecule attached to a nucleotide of the polynucleotide by means of a linker arm, and (ii) a background-reducing reagent which chemically modifies the intercalating molecule when the probe to which it is attached is single-stranded; and (b) detecting a property change resulting from the intercalation of the intercalating molecule into a target-probe hybrid, thereby detecting the target polynucleotide. The intercalating molecule which is part of the polynucleotide probe induces a change in a property, in either the probe, the target polynucleotide or a target-probe hybrid. The property change can be detected, for example, by means of a generated signal which can be identified or quantified. The present invention can be employed in a heterogeneous (two step or two phase) assay using a support to immobilize the target or probe, and a washing step, and in a homogeneous (one step or one phase) assay using a hybridization solution. Also provided are a composition and a nucleic acid hybridization kit useful for detecting the presence of a target polynucleotide in a sample.

Abstract translation: 本发明提供了一种用于检测样品中靶多核苷酸的存在的方法。 该方法包括以下步骤：（a）在杂交条件下使样品与（i）能够与靶多核苷酸杂交的单链多核苷酸探针接触，并且包含多核苷酸和至少一个与多核苷酸的核苷酸连接的插入分子， 连接臂的手段，和（ii）当与其连接的探针单链时化学修饰插入分子的背景降低试剂; 和（b）检测由插入分子插入到靶 - 探针杂交体中产生的性质变化，从而检测靶多核苷酸。 作为多核苷酸探针的一部分的插入分子在探针，靶多核苷酸或靶 - 探针杂交体中诱导性质的变化。 可以例如通过可以被识别或量化的生成信号来检测属性变化。 本发明可以在使用支持物固定靶或探针，洗涤步骤和使用杂交溶液的均相（一步或一相）测定）的异质（两步或两相）测定中使用。 还提供了可用于检测样品中靶多核苷酸的存在的组合物和核酸杂交试剂盒。

公开(公告)号：US4849505A

公开(公告)日：1989-07-18

申请号：US43572

申请日：1987-04-28

Applicant: Jannis G. Stavrianopoulos

Inventor： Jannis G. Stavrianopoulos

IPC: A61K51/04 ,  A61K51/10 ,  G01N33/533 ,  G01N33/553

CPC classification number: A61K51/1093 ,  A61K51/0497 ,  G01N33/533 ,  G01N33/553

Abstract: A detectable molecule of the formulaA.sup.3 --(--X--R.sup.1 --E--Det.sup.b).sub.mwhere A.sup.3 is A.sup.2 or a polymer, where A.sup.3 has at least one modifiable reactive group selected from the group consisting of amino, hydroxy, cis OH, halides, aryl, imidazoyl, carbonyl, carboxy, thiol or a residue comprising an activated carbon; --X-- is selected from the group consisting of ##STR1## --R.sup.1 -- is ##STR2## or a C.sub.1 -C.sub.10 branched or unbranched alkyl or aralkyl, which may be substituted by --OH; --Y-- is a direct bond to --E--, or --Y-- is --E--R.sup.2 -- where R.sup.2 is a C.sub.1 -C.sub.10 branched or unbranched alkyl; Z.sub.a is chlorine, bromine or iodine; E is O, NH or an acyclic divalent sulfur atom; Det.sup.b is a chemical moiety capable of being detected, preferably comprising biotin or a metal chelator of the formula: ##STR3## or the 4-hydroxy or acyloxy derivatives thereof, where R.sup.3 is C.sub.1 -C.sub.4 alkyl or CH.sub.2 COOM, M is the same or different and selected from the group consisting of hydrogen, a metal or non-metal cation or is C.sub.1 -C.sub.10 alkyl, aryl or aralkyl; and m is an integer from 1 to the total number of modified reactive groups on A.sup.3. The detectable molecules are useful in in vitro or in vivo assays or therapy.

Abstract translation: 式A3-（ - X-R1-E-Detb）m的可检测分子，其中A3是A2或聚合物，其中A3具有选自氨基，羟基，顺式OH，卤化物中的至少一个可修饰的反应性基团 ，芳基，咪唑基，羰基，羧基，硫醇或包含活性炭的残基; -X-选自 + TR -R1-是或可以被-OH取代的C1-C10支链或非支链烷基或芳烷基 ; -Y-是与-E-或-Y-是-E-R 2 - 的直接键合，其中R 2是C 1 -C 10支链或非支链烷基; Za是氯，溴或碘; E是O，NH或无环二价硫原子; Detb是能够被检测的化学部分，优选包含下式的生物素或金属螯合剂：其中R 3是C 1 -C 4烷基或CH 2 COOM，M相同或不同 并且选自氢，金属或非金属阳离子或是C 1 -C 10烷基，芳基或芳烷基; 并且m是从1到A3上修饰的反应性基团的总数的整数。 可检测的分子可用于体外或体内测定或治疗。

公开(公告)号：US4849208A

公开(公告)日：1989-07-18

申请号：US43577

申请日：1987-04-28

Applicant: Jannis G. Stavrianopoulos

Inventor： Jannis G. Stavrianopoulos

IPC: A61K51/04 ,  A61K51/10 ,  G01N33/533 ,  G01N33/553

CPC classification number: A61K51/0497 ,  A61K51/1093 ,  G01N33/533 ,  G01N33/553

Abstract: A detectable molecule of the formulaA.sup.3 --(--X--R.sup.1 --E--Det.sup.b).sub.mwherein A.sup.3 is A.sup.2 or a polymer, where A.sup.3 has at least one modifiable reactive group selected from the group consisting of amino, hydroxy, cis .OH, halides, aryl, imidazoly, carbonyl, carboxy, thiol or a residue comprising an activated carbon; --X-- is selected from the group consisting of ##STR1## a C.sub.1 -C.sub.10 branched or unbranched alkyl or aralkyl, which may be substituted by --OH; --Y-- is a direct bond to --E--, or --Y-- is --E--R.sup.2 -- where R.sup.2 is a C.sub.1 -C.sub.10 branched or unbranched alkyl; Z.sub.a is chlorine, bromine or iodine; E is O, NH or an acyclic divalent sulfur atom; Det.sup.b is a chemical moiety capable of being detected, preferably comprising biotin or a metal chelator of the formula: ##STR2## or the 4-hydroxy or acyloxy derivative thereof, where R.sup.3 is C.sub.1 -C.sub.4 alkyl or CH.sub.2 COOM, M is the same or different and selected from the group consisting of hydrogen, a metal or non-metal cation or is C.sub.1 -C.sub.10 alkyl, aryl or aralkyl; and mm is an integer from 1 to the total number of modified reactive groups on A.sup.3. The detectable molecules are useful in in vitro or in vivo assays or therapy.

Abstract translation: 式A3-（-X-R1-E-Detb）m的可检测分子，其中A3是A2或聚合物，其中A3具有至少一个选自氨基，羟基，顺式-OH， 卤化物，芳基，咪唑，羰基，羧基，硫醇或包含活性炭的残基; -X-选自可以被-OH取代的C1-C10支链或非支链烷基或芳烷基的组合 OCONH + TR -Y-是与-E-或-Y-是-E-R 2 - 的直接键合，其中R 2是C 1 -C 10支链或非支链烷基; Za是氯，溴或碘; E是O，NH或无环二价硫原子; Detb是能够被检测的化学部分，优选包含下式的生物素或金属螯合剂：其中R 3是C 1 -C 4烷基或CH 2 COOM，M相同或不同 并且选自氢，金属或非金属阳离子或是C 1 -C 10烷基，芳基或芳烷基; 并且mm是从1到A3上修饰的反应性基团总数的整数。 可检测的分子可用于体外或体内测定或治疗。

公开(公告)号：US08389702B2

公开(公告)日：2013-03-05

申请号：US12803355

申请日：2010-06-24

Applicant: Elazar Rabbani ,  Jannis G. Stavrianopoulos ,  James J. Donegan

Inventor： Elazar Rabbani ,  Jannis G. Stavrianopoulos ,  James J. Donegan

IPC: C07H21/00 ,  C07H21/02 ,  C07H19/02 ,  C12Q1/68 ,  C12P19/34

CPC classification number: C07H21/04

Abstract: The present invention provides multisignal labeling reagents and these are useful in a number of biochemical applications, including the manufacture of biomolecular probes and their use in detecting or amplifying analyte-specific moieties.

Abstract translation: 本发明提供多信号标记试剂，并且它们可用于许多生化应用，包括生物分子探针的制备及其在检测或扩增分析物特异性部分中的用途。

公开(公告)号：US08133989B2

公开(公告)日：2012-03-13

申请号：US10307108

申请日：2002-11-27

Applicant: Elazar Rabbani ,  Jannis G. Stavrianopoulos ,  James J. Donegan ,  Jack Coleman ,  Marleen Walner

Inventor： Elazar Rabbani ,  Jannis G. Stavrianopoulos ,  James J. Donegan ,  Jack Coleman ,  Marleen Walner

IPC: C07H21/04 ,  C12P19/34

CPC classification number: C12Q1/6869 ,  C12P19/34 ,  C12Q1/6844 ,  C12Q1/6853 ,  C12Q2531/119 ,  C12Q2525/301 ,  C12Q2525/161

Abstract: This invention provides novel processes for amplifying nucleic acid sequences of interest, including linear and non-linear amplification. In linear amplification, a single initial primer or nucleic acid construct is utilized to carry out the amplification process. In non-linear amplification, a first initial primer or nucleic acid construct is employed with a subsequent initial primer or nucleic acid construct. In other non-linear amplification processes provided by this invention, a first initial primer or nucleic acid construct is deployed with a second initial primer or nucleic acid construct to amplify the specific nucleic acid sequence of interest and its complement that are provided. A singular primer or a singular nucleic acid construct capable of non-linear amplification can also be used to carry out non-linear amplification in accordance with this invention. Post-termination labeling process for nucleic acid sequencing is also disclosed in this invention that is based upon the detection of tagged molecules that are covalently bound to chemically reactive groups provided for chain terminators. A process for producing nucleic acid sequences having decreased thermodynamic stability to complementary sequences is also provided and achieved by this invention. Unique nucleic acid polymers are also disclosed and provided in addition to other novel compositions, kits and the like.

Abstract translation: 本发明提供用于扩增感兴趣的核酸序列的新方法，包括线性和非线性扩增。 在线性扩增中，使用单个初始引物或核酸构建体进行扩增过程。 在非线性扩增中，使用第一初始引物或核酸构建体与随后的初始引物或核酸构建体。 在本发明提供的其它非线性扩增方法中，用第二初始引物或核酸构建物展开第一个初始引物或核酸构建体，以扩增所提供的特异性核酸序列及其互补序列。 也可以使用能够进行非线性扩增的单一引物或单个核酸构建体来进行本发明的非线性扩增。 用于核酸测序的终止后标记方法也在本发明中公开，其基于共价结合到为链终止剂提供的化学反应性基团的标记分子的检测。 通过本发明提供并实现了对互补序列产生具有降低的热力学稳定性的核酸序列的方法。 除了其它新型组合物，试剂盒等之外，还公开并提供了独特的核酸聚合物。

公开(公告)号：US20110098441A1

公开(公告)日：2011-04-28

申请号：US10763104

申请日：2004-01-22

Applicant: Jannis G. Stavrianopoulos ,  Flazar Rabbani

Inventor： Jannis G. Stavrianopoulos ,  Flazar Rabbani

IPC: C07K5/103 ,  C07H19/10 ,  C07K9/00 ,  C07K19/00 ,  C07H21/00 ,  C07K16/46 ,  C08B37/00 ,  C07D403/06 ,  C07D311/82 ,  C07D491/16

CPC classification number: C07H21/00 ,  C09B23/02 ,  C09B47/00 ,  C09B57/00 ,  Y10S436/80

Abstract: This invention provides for labeling reagents, labeled targets and processes for preparing labeling reagents. The labeling reagents can take the form of cyanine dyes, xanthene dyes, porphyrin dyes, coumarin dyes or composite dyes. These labeling reagents are useful for labeling probes or targets, including nucleic acids and proteins. These reagents can be usefully applied to protein and nucleic acid probe based assays. They are also applicable to real-time detection processes.

Abstract translation: 本发明提供标记试剂，标记的靶标和制备标记试剂的方法。 标记试剂可以采用花青染料，呫吨染料，卟啉染料，香豆素染料或复合染料的形式。 这些标记试剂可用于标记探针或靶标，包括核酸和蛋白质。 这些试剂可用于基于蛋白质和基于核酸探针的测定。 它们也适用于实时检测过程。

公开(公告)号：US20110097791A1

公开(公告)日：2011-04-28

申请号：US10260031

申请日：2003-06-06

Applicant: Dean L. Engelhardt ,  Jannis G. Stavrianopoulos ,  Elazar Rabbani ,  James J. Donegan

Inventor： Dean L. Engelhardt ,  Jannis G. Stavrianopoulos ,  Elazar Rabbani ,  James J. Donegan

IPC: C12N15/63

CPC classification number: C12N15/10 ,  C12N15/79 ,  C12P19/34 ,  C12Q1/6844 ,  C12Q1/6853 ,  C12Q1/6858 ,  C12Q1/686 ,  C12Q1/6865 ,  C12Q2525/203 ,  C12Q2521/119

Abstract: This invention provides inter alia an in vitro process for producing multiple specific nucleic acid copies in which the copies are produced under isostatic conditions, e.g., temperature, buffer and ionic strength, and independently of any requirement for introducing an intermediate structure for producing the copies. In other aspects, the invention provides in vitro processes for producing multiple specific nucleic acid copies in which the products are substantially free of any primer-coded sequences, such sequences having been substantially or all removed from the product to regenerate a primer binding site, thereby allowing new priming events to occur and multiple nucleic acid copies to be produced. This invention further provides a promoter-independent non-naturally occurring nucleic acid construct that produces a nucleic acid copy or copies without using or relying on any gene product that may be coded by the nucleic acid construct. Another aspect of this invention concerns a protein-nucleic acid construct in the form of a conjugate linked variously, e.g., covalent linkage, complementary nucleic acid base-pairing, nucleic acid binding proteins, or ligand receptor binding. Further disclosed in this invention is an in vivo process for producing a specific nucleic acid in which such a protein-nucleic acid construct conjugate is introduced into a cell. A still further aspect of the invention relates to a construct comprising a host promoter, second promoter and DNA sequence uniquely located on the construct. The host transcribes a sequence in the construct coding for a different RNA polymerase which after translation is capable of recognizing its cognate promoter and transcribing from a DNA sequence of interest in the construct with the cognate promoter oriented such that it does not promote transcription from the construct of the different RNA polymerase.

Abstract translation: 本发明特别提供用于生产多种特异性核酸拷贝的体外方法，其中拷贝在等静压条件下，例如温度，缓冲液和离子强度下产生，并且独立于引入用于产生拷贝的中间结构的任何要求。 在其它方面，本发明提供了用于产生多种特异性核酸拷贝的体外方法，其中产物基本上不含任何引物编码的序列，这些序列基本上或全部从产物中除去以再生引物结合位点，从而 允许发生新的引发事件并产生多个核酸拷贝。 本发明进一步提供了一种与天然存在的启动子相关的非天然存在的核酸构建体，其不使用或依赖于可由核酸构建体编码的任何基因产物产生核酸拷贝或拷贝。 本发明的另一方面涉及以不同方式连接的缀合物形式的蛋白质 - 核酸构建体，例如共价连接，互补核酸碱基配对，核酸结合蛋白或配体受体结合。 在本发明中进一步公开的是用于生产其中将这种蛋白质 - 核酸构建体缀合物引入细胞的特异性核酸的体内方法。 本发明的另一方面涉及包含宿主启动子，第二启动子和唯一位于构建体上的DNA序列的构建体。 宿主转录编码不同RNA聚合酶的构建体中的序列，其在翻译后能够识别其同源启动子并且从构建体中感兴趣的DNA序列转录成同源启动子定向，使得其不促进构建体的转录 的不同RNA聚合酶。

公开(公告)号：US20110028692A1

公开(公告)日：2011-02-03

申请号：US12803354

申请日：2010-06-24

Applicant: Elazar Rabbani ,  Jannis G. Stavrianopoulos ,  James J. Donegan

Inventor： Elazar Rabbani ,  Jannis G. Stavrianopoulos ,  James J. Donegan

IPC: C07H21/00 ,  C07K14/765 ,  C07K14/00

CPC classification number: C07H21/04

Abstract: The present invention provides multisignal labeling reagents and these are useful in a number of biochemical applications, including the manufacture of biomolecular probes and their use in detecting or amplifying analyte-specific moieties.

Abstract translation: 本发明提供多信号标记试剂，并且它们可用于许多生化应用，包括生物分子探针的制备及其在检测或扩增分析物特异性部分中的用途。