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21. 发明授权

US08071794B2 Long wavelength thiol-reactive fluorophores 有权
标题翻译：长波长硫醇反应性荧光团
公开(公告)号：US08071794B2
公开(公告)日：2011-12-06
申请号：US12489145
申请日：2009-06-22
申请人： J. Bruce Pitner , Douglas B. Sherman , Joseph Thomas
发明人： J. Bruce Pitner , Douglas B. Sherman , Joseph Thomas
IPC分类号： C07D413/06 , A61K31/538 , G01N33/48
CPC分类号： C09B23/10 , C07D207/02 , C09B23/105 , G01N33/582 , G01N33/66
摘要： Reactive fluorescent dyes compositions and methods of using same are disclosed. Squaraine nucleus, Nile Red nucleus, benzodioxazole nucleus, coumarin nucleus or aza coumarin nucleus dyes are disclosed having thiol-reactive groups. Squaraine nucleus, Nile Red nucleus, benzodioxazole nucleus, coumarin nucleus or aza coumarin nucleus dyes are disclosed that exhibit a fluorescence emission of at least about 575 nm. Biosensors are disclosed having a binding protein and a squaraine nucleus, Nile Red nucleus, benzodioxazole nucleus, coumarin nucleus or aza coumarin nucleus.
摘要翻译：公开了活性荧光染料组合物及其使用方法。揭示了具有硫醇反应性基团的方钠核，尼罗红核，苯并恶唑核，香豆素核或氮杂香豆素核染料。披露了方解子核，尼罗红核，苯并恶唑核，香豆素核或者aza香豆素核染料，其表现出至少约575nm的荧光发射。公开了具有结合蛋白和方酸核，尼罗红核，苯并恶唑核，香豆素核或者aza香豆素核的生物传感器。

22. 发明授权

US07316909B2 Binding protein as biosensors 失效
标题翻译：结合蛋白作为生物传感器
公开(公告)号：US07316909B2
公开(公告)日：2008-01-08
申请号：US11424892
申请日：2006-06-19
申请人： J. Bruce Pitner , Jason E. Gestwicki , Zachary A. Pfieffer , Helen V. Hsieh
发明人： J. Bruce Pitner , Jason E. Gestwicki , Zachary A. Pfieffer , Helen V. Hsieh
IPC分类号： G01N33/53
CPC分类号： G01N33/66 , G01N33/54373
摘要： The invention is directed to compositions of mutated binding proteins containing thiol groups for coupling to sensor surfaces, analyte biosensor devices derived there from, and methods of their use as analyte biosensors both in vitro and in vivo.
摘要翻译：本发明涉及含有用于偶联至传感器表面的硫醇基团的衍生的结合蛋白的组合物，从其衍生的分析物生物传感器装置及其在体外和体内作为分析物生物传感器的方法。

23. 发明授权

US06900030B2 Device for monitoring cells 有权
标题翻译：用于监控单元的设备
公开(公告)号：US06900030B2
公开(公告)日：2005-05-31
申请号：US10116777
申请日：2002-04-04
申请人： J. Bruce Pitner , John Jacob Hemperly , Richard D. Guarino , Magdalena Wodnicka , David T. Stitt , Gregory J. Burrell , Timothy G. Foley, Jr. , Patrick Shawn Beaty
发明人： J. Bruce Pitner , John Jacob Hemperly , Richard D. Guarino , Magdalena Wodnicka , David T. Stitt , Gregory J. Burrell , Timothy G. Foley, Jr. , Patrick Shawn Beaty
IPC分类号： C12Q1/04 , C12Q1/18
CPC分类号： C12Q1/18 , C12Q1/04 , C12Q2304/44 , G06F19/00 , Y02A90/26
摘要： The present invention relates to methods for detection and evaluation of metabolic activity of eukaryotic and/or prokaryotic cells based upon their ability to consume dissolved oxygen. The methods utilize a luminescence detection system which makes use of the sensitivity of the luminescent emission of certain compounds to the presence of oxygen, which quenches (diminishes) the compound's luminescent emission in a concentration dependent manner. Respiring eukaryotic and/or prokaryotic cells will affect the oxygen concentration of a liquid medium in which they are immersed. Thus, this invention provides a convenient system to gather information on the presence, identification, quantification and cytotoxic activity of eukaryotic and/or prokaryotic cells by determining their effect on the oxygen concentration of the media in which they are present.
摘要翻译：本发明涉及基于其消耗溶解氧的能力来检测和评价真核细胞和/或原核细胞的代谢活性的方法。该方法利用了发光检测系统，其利用某些化合物的发光发射对氧的存在的敏感性，其以浓度依赖性方式猝灭（减少）化合物的发光发射。呼吸真核细胞和/或原核细胞将影响浸入其中的液体培养基的氧浓度。因此，本发明提供了一种方便的系统，用于通过确定它们对存在的介质的氧浓度的影响来收集关于真核细胞和/或原核细胞的存在，鉴定，定量和细胞毒活性的信息。

24. 发明授权

US06379888B1 Universal probes and methods for detection of nucleic acids 失效
标题翻译：用于检测核酸的通用探针和方法
公开(公告)号：US06379888B1
公开(公告)日：2002-04-30
申请号：US09406074
申请日：1999-09-27
申请人： James G. Nadeau , C. Preston Linn , J. Bruce Pitner , Cheryl H. Dean , G. Terrance Walker
发明人： James G. Nadeau , C. Preston Linn , J. Bruce Pitner , Cheryl H. Dean , G. Terrance Walker
IPC分类号： C12Q168
CPC分类号： C12Q1/6818 , Y10T436/143333 , C12Q2563/137 , C12Q2537/1373
摘要： Signal primers are employed for detection of nucleic acid target sequences by fluorescence quenching mechanisms. The signal primer comprises a first and a second oligonucleotide and is partially single-stranded and partially double-stranded. In the presence of target, the second oligonucleotide of the signal primer is displaced from the first and a conformational change in a reporter probe occurs which changes the distance between the members of a donor/quencher dye pair linked to the reporter probe. The change in proximity between the dyes causes an increase or a decrease in fluorescence quenching, which is detected as an indication of the presence of the target sequence.
摘要翻译：信号引物用于通过荧光猝灭机制检测核酸靶序列。信号引物包含第一和第二寡核苷酸并且是部分单链和部分双链的。在靶的存在下，信号引物的第二寡核苷酸从第一个位点移位，并且发生报告物探针中的构象变化，其改变与报道探针连接的供体/猝灭剂染料对的成员之间的距离。染料之间接近度的变化导致荧光淬灭的增加或减少，其被检测为靶序列存在的指示。

25. 发明授权

US06261784B1 Detection of nucleic acids by strand displacement 有权
标题翻译：通过链置换检测核酸
公开(公告)号：US06261784B1
公开(公告)日：2001-07-17
申请号：US09599164
申请日：2000-06-22
申请人： James G. Nadeau , Helen V. Hsieh , J. Bruce Pitner , C. Preston Linn
发明人： James G. Nadeau , Helen V. Hsieh , J. Bruce Pitner , C. Preston Linn
IPC分类号： C12Q168
CPC分类号： C12Q1/6818 , C12Q2537/1373 , C12Q2531/143 , C12Q2531/119
摘要： Detector nucleic acids are employed for detection of nucleic acid target sequences by fluorescence quenching mechanisms. The detector nucleic acid comprises at least two oligonucleotides and is partially single-stranded and partially double-stranded. One of the two dyes of a donor/acceptor dye pair is linked to the first oligonucleotide and the other is linked to a second oligonucleotide such that they are in close spatial proximity when the first and second oligonucleotides are base-paired and donor fluorescence is quenched. A single second oligonucleotide may be hybridized to the first oligonucleotide or multiple second oligonucleotides may be hybridized to the first oligonucleotide and to each other, forming a junction structure comprising multiple donor/acceptor dye pairs. The detector oligonucleotide retains its partially single-stranded and partially double-stranded conformation in the absence of target. In the presence of target, however, the second oligonucleotide(s) of the detector nucleic acid is/are completely or partially displaced from the first, increasing the distance between the donor and acceptor dyes and causing a change in fluorescence which may be detected as an indication of the presence of the target sequence.
摘要翻译：检测器核酸用于通过荧光猝灭机制检测核酸靶序列。检测器核酸包含至少两个寡核苷酸并且是部分单链和部分双链的。供体/受体染料对的两种染料之一与第一寡核苷酸连接，另一种与第二寡核苷酸连接，使得当第一和第二寡核苷酸是碱基配对并且供体荧光淬灭时它们处于紧密的空间接近。单个第二寡核苷酸可以与第一寡核苷酸杂交，或者多个第二寡核苷酸可以与第一寡核苷酸和彼此杂交，形成包含多个供体/受体染料对的连接结构。检测器寡核苷酸在不存在目标时保留其部分单链和部分双链构象。然而，在靶存在下，检测器核酸的第二寡核苷酸与第一寡核苷酸完全或部分位移，从而增加供体和受体染料之间的距离并引起荧光的变化，其可被检测为指示靶序列的存在。

26. 发明授权

US6130047A Detection of nucleic acids by fluorescence quenching 失效
标题翻译：通过荧光猝灭检测核酸
公开(公告)号：US6130047A
公开(公告)日：2000-10-10
申请号：US235583
申请日：1999-01-22
申请人： James G. Nadeau , Helen V. Hsieh , J. Bruce Pitner , C. Preston Linn
发明人： James G. Nadeau , Helen V. Hsieh , J. Bruce Pitner , C. Preston Linn
IPC分类号： G01N21/64 , C07H21/00 , C12N15/09 , C12Q1/68 , G01N33/542 , G01N33/566 , C07H21/04 , C12P19/34
CPC分类号： C12Q1/6818
摘要： Detector nucleic acids are employed for detection of nucleic acid target sequences by fluorescence quenching mechanisms. The detector nucleic acid comprises at least two oligonucleotides and is partially single-stranded and partially double-stranded. One of the two dyes of a donor/acceptor dye pair is linked to the first oligonucleotide and the other is linked to a second oligonucleotide such that they are in close spatial proximity when the first and second oligonucleotides are base-paired and donor fluorescence is quenched. A single second oligonucleotide may be hybridized to the first oligonucleotide or multiple second oligonucleotides may be hybridized to the first oligonucleotide and to each other, forming a junction structure comprising multiple donor/acceptor dye pairs. The detector oligonucleotide retains its partially single-stranded and partially double-stranded conformation in the absence of target. In the presence of target, however, the second oligonucleotide(s) of the detector nucleic acid is/are completely or partially displaced from the first, increasing the distance between the donor and acceptor dyes and causing a change in fluorescence which may be detected as an indication of the presence of the target sequence.
摘要翻译：检测器核酸用于通过荧光猝灭机制检测核酸靶序列。检测器核酸包含至少两个寡核苷酸并且是部分单链和部分双链的。供体/受体染料对的两种染料之一与第一寡核苷酸连接，另一种与第二寡核苷酸连接，使得当第一和第二寡核苷酸是碱基配对并且供体荧光淬灭时它们处于紧密的空间接近。单个第二寡核苷酸可以与第一寡核苷酸杂交，或者多个第二寡核苷酸可以与第一寡核苷酸和彼此杂交，形成包含多个供体/受体染料对的连接结构。检测器寡核苷酸在不存在目标时保留其部分单链和部分双链构象。然而，在靶存在下，检测器核酸的第二寡核苷酸与第一寡核苷酸完全或部分位移，从而增加供体和受体染料之间的距离并引起荧光的变化，其可被检测为指示靶序列的存在。

27. 发明授权

US6054279A Detection of nucleic acids by fluorescence quenching 失效
标题翻译：通过荧光猝灭检测核酸
公开(公告)号：US6054279A
公开(公告)日：2000-04-25
申请号：US120916
申请日：1998-07-20
申请人： James G. Nadeau , J. Bruce Pitner , James L. Schram , C. Preston Linn , Glenn P. Vonk , G. Terrance Walker
发明人： James G. Nadeau , J. Bruce Pitner , James L. Schram , C. Preston Linn , Glenn P. Vonk , G. Terrance Walker
IPC分类号： G01N21/64 , C07H21/00 , C12N15/09 , C12Q1/68 , G01N33/58 , C21Q1/68
CPC分类号： C12Q1/6818 , C12Q1/6825
摘要： Single-stranded signal primers are modified by linkage to two dyes which form a donor/acceptor dye pair. The two dyes are positioned in sufficiently close spatial proximity on the signal primer that the fluorescence of the first dye is quenched by the second dye. The signal primer may further comprise a restriction endonuclease recognition site (RERS) between the two dyes. As the signal primer is initially single-stranded and remains single-stranded in the absence of target, the restriction endonuclease recognition site is not cleavable or nickable by the restriction endonuclease. In the presence of target, however, signal primer and the restriction endonuclease recognition site are rendered double-stranded and cleavable or nickable by the restriction endonuclease. Cleavage or nicking separates the two dyes and a change in fluorescence due to decreased quenching is detected as an indication of the presence of the target sequence or of target sequence amplification.
摘要翻译：通过连接到形成供体/受体染料对的两种染料来修饰单链信号引物。这两种染料位于信号引物足够接近的空间上，第一种染料的荧光被第二种染料淬灭。信号引物还可以包含两种染料之间的限制性内切核酸酶识别位点（RERS）。由于信号引物最初是单链的，并且在不存在靶标的情况下保持单链，限制性内切核酸酶识别位点不受限制性内切核酸酶的切割或切割。然而，在靶的存在下，信号引物和限制性内切核酸酶识别位点被限制性内切核酸酶双链和切割或切割。切割或切口分离两种染料，并且由于猝灭降低引起的荧光变化被检测为目标序列或靶序列扩增的存在的指示。

28. 发明授权

US5958700A Detection of nucleic acids by fluorescence quenching 失效
标题翻译：通过荧光猝灭检测核酸
公开(公告)号：US5958700A
公开(公告)日：1999-09-28
申请号：US237510
申请日：1999-01-26
申请人： James G. Nadeau , J. Bruce Pitner , C. Preston Linn , James L. Schram
发明人： James G. Nadeau , J. Bruce Pitner , C. Preston Linn , James L. Schram
IPC分类号： G01N33/50 , C12N15/09 , C12Q1/68 , G01N33/566 , C07H21/04 , C12P19/34
CPC分类号： C12Q1/6827 , C12Q1/6818 , C12Q1/6858
摘要： A detector oligonucleotide having a sequence which forms an intramolecularly base-paired secondary structure is described for use in detecting nucleic acid target sequences and target sequence amplification. The detector oligonucleotide is further modified by linkage to two dyes which form a donor/acceptor dye pair. The two dyes are positioned on the detector oligonucleotide such that they are in close spatial proximity in the base-paired, folded secondary structure, thereby causing quenching of donor fluorescence. The detector oligonucleotide may optionally further comprise a restriction endonuclease recognition site (RERS) which remains partially or entirely single-stranded in the base-paired secondary structure. The RERS is flanked by the two dyes. In the presence of target, the base-paired secondary structure is unfolded or linearized, increasing the distance between the donor and acceptor dyes and causing a change in fluorescence of the donor and/or the acceptor. If an RERS is present, it is rendered double-stranded in the presence of target, allowing cleavage or nicking by a restriction endonuclease and separation of the two dyes onto separate nucleic acid fragments. This may further contribute to the magnitude of the change in fluorescence.
摘要翻译：描述了具有形成分子内碱基配对二级结构的序列的检测器寡核苷酸用于检测核酸靶序列和靶序列扩增。通过连接到形成供体/受体染料对的两种染料进一步修饰检测器寡核苷酸。两种染料位于检测器寡核苷酸上，使得它们在碱基配对的折叠的二级结构中处于紧密的空间接近处，由此引起供体荧光的猝灭。检测器寡核苷酸可以任选地进一步包含限制性内切核酸酶识别位点（RERS），所述限制性内切核酸酶识别位点在碱基配对二级结构中部分或全部保持单链。 RERS侧面有两种染料。在靶的存在下，碱基配对的二级结构被展开或线性化，增加供体和受体染料之间的距离并引起供体和/或受体的荧光变化。如果存在RERS，则在靶的存在下使其成为双链，允许通过限制性内切核酸酶切割或切割并将两种染料分离到单独的核酸片段上。这可能进一步有助于荧光变化的大小。

29. 发明授权

US5691145A Detection of nucleic acids using G-quartets 失效
标题翻译：使用G四重奏检测核酸
公开(公告)号：US5691145A
公开(公告)日：1997-11-25
申请号：US703755
申请日：1996-08-27
申请人： J. Bruce Pitner , Glenn P. Vonk , James G. Nadeau
发明人： J. Bruce Pitner , Glenn P. Vonk , James G. Nadeau
IPC分类号： G01N33/58 , C07H21/04 , C12N15/09 , C12Q1/68 , G01N21/78 , C07H21/02 , C12P19/34
CPC分类号： C12Q1/6818 , Y10S436/80 , Y10S436/805 , Y10T436/143333
摘要： Oligonucleotides which form G-quartet structures have been found to be useful in fluorescence assays to detect a selected nucleic acid sequence. When one end of the oligonucleotide is labeled with a donor fluorophore and the other end is labeled with an acceptor dye, the folding of the molecule in the G-quartet structure brings the donor-acceptor pair into close proximity, allowing an interaction between the two labels which results in quenching of donor fluorescence or a change in other fluorescence properties which are the result of the interaction of two dyes in close proximity. The G-quartet structure unfolds upon hybridization to its complementary sequence, increasing the distance between the two dye labels. This results in decreased donor quenching or a change in another proximity-related fluorescence parameter. The associated increase in donor fluorescence intensity or the change in another fluorescence parameter may be monitored as an indication of the presence of a selected nucleic acid sequence. Alternatively, in some cases a decrease in acceptor fluorescence may be monitored as an indication of the presence of the selected nucleic acid sequence when the acceptor is also a fluorophore.
摘要翻译：已经发现形成G-四重结构的寡核苷酸可用于荧光测定以检测所选择的核酸序列。当寡核苷酸的一端用供体荧光团标记，另一端用受体染料标记时，分子在G四重结构中的折叠使供体 - 受体对紧密接近，允许两者之间的相互作用导致供体荧光猝灭的标记或其他荧光性质的变化，这两种染料是两种染料相互作用的结果。 G-四重结构在与其互补序列杂交时展开，增加两个染料标记之间的距离。这导致供体猝灭减少或另一邻近相关荧光参数的变化。可以监测供体荧光强度的相关增加或另一荧光参数的变化作为选择的核酸序列的存在的指示。或者，在一些情况下，当受体也是荧光团时，可以监测受体荧光的降低作为选择的核酸序列的存在的指示。

30. 发明授权

US5393514A Fluorescent pH indicators 失效
公开(公告)号：US5393514A
公开(公告)日：1995-02-28
申请号：US162554
申请日：1993-12-06
申请人： J. Bruce Pitner , Randal A. Hoke
发明人： J. Bruce Pitner , Randal A. Hoke
IPC分类号： C09B11/28 , C07D311/82 , C09B11/24 , C09K11/06 , G01N21/80 , G01N31/22 , A61K49/00 , A61K31/35
CPC分类号： C09B11/24 , C07D311/82 , G01N31/221
摘要： Fluorescent pH indicator compounds which exhibit an increase in fluorescence intensity with decreasing pH. The indicators are derivatives of rhodamine and sulforhodamine type dyes and are particularly useful for measuring pH in acidic environments such as carbon dioxide production by microorganisms and pH of certain acidic intracellular compartments.

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