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公开(公告)号：US06420115B1
公开(公告)日：2002-07-16
申请号：US09613263
申请日：2000-07-10
IPC分类号： C12Q168
CPC分类号： C12Q1/6839 , C12Q1/6816 , C12Q1/6827 , C12Q2563/137 , C12Q2563/107 , C12Q2563/173
摘要： Triplex complexes contain a single-stranded probe bound to a double-stranded nucleic acid target, in which the probe includes a heteropolymeric nucleic acid or a heteropolymeric nucleic acid analog. All base triplets of the complex are members selected from the group consisting of A-T-A, T-A-T, U-A-T, T-A-U, A-U-A, U-A-U, G-C-G and C-G-C. A cation-facilitated assay includes detecting the presence of such triplex complexes to determine the degree of complementarity between the probe and target sequence. The assay preferably detects a change in fluorescent intensity of a label as a function of binding affinity between the probe and target. The label can be covalently tethered to the probe or to the target, or can be an intercalating fluorophore in the reaction medium.
摘要翻译： 三重配合物含有与双链核酸靶结合的单链探针，其中探针包括杂聚核酸或异源核酸类似物。 复合体的所有基本三元组是从由A-T-A，T-A-T，U-A-T，T-A-U，A-U-A，U-A-U，G-C-G和C-G-C组成的组中选出的成员。 阳离子促进测定法包括检测这种三重配合物的存在以确定探针和靶序列之间的互补程度。 该测定优选检测作为探针和靶标之间的结合亲合力的函数的标记物的荧光强度的变化。 标记可以共价连接到探针或靶上，或者可以是反应介质中的插层荧光团。
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2.发明授权Fluorescent intensity method for assaying binding between proteins or peptides 失效用于测定蛋白质或肽之间的结合的荧光强度法公开(公告)号：US06645733B1
公开(公告)日：2003-11-11
申请号：US09344525
申请日：1999-06-25
申请人： Jasmine I. Daksis , Glen H. Erikson
发明人： Jasmine I. Daksis , Glen H. Erikson
IPC分类号： G01N2100
CPC分类号： G01N33/542 , Y10S435/81
摘要： A method for assaying specific binding between a fluorophore-labeled probe and an unlabeled target is provided. The method includes detecting a quenching effect on fluorescence emitted by the fluorophore-labeled probe resulting from binding. The method is conducted without separating complexes of the target and probe from the free target and free probe prior to quenching effect detecting, and without providing a signal quenching agent to quench fluorescent light. Preferably, the probe and target are amino acid-containing compounds, such as proteins. The method can be used for a variety of applications, including screening for drug candidates having optimum binding properties, and quantifying and classifying the binding characteristics between peptide-containing compounds. The method is more sensitive than conventional assays, enabling the analysis of minute samples and low affinity binding interactions between receptors and ligands that are below the detection limits of conventional technology.
摘要翻译： 提供了用于测定荧光团标记的探针和未标记靶之间特异性结合的方法。 该方法包括检测由结合产生的由荧光团标记的探针发射的荧光的淬灭效应。 在淬灭效应检测之前不进行目标物和探针的混合物与游离靶和游离探针的分离，而不提供信号淬灭剂来淬灭荧光。 优选地，探针和靶是含有氨基酸的化合物，例如蛋白质。 该方法可用于各种应用，包括筛选具有最佳结合特性的候选药物，以及量化和分类含肽化合物之间的结合特征。 该方法比常规测定更敏感，能够分析低于常规技术检测限的受体和配体之间的微小样品和低亲和力结合相互作用。
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3.发明授权公开(公告)号：US06294333B1
公开(公告)日：2001-09-25
申请号：US09224505
申请日：1998-12-31
申请人： Jasmine I. Daksis , Glen H. Erikson
发明人： Jasmine I. Daksis , Glen H. Erikson
IPC分类号： C12Q168
CPC分类号： C12Q1/6804 , C12Q1/6818 , G01N33/542 , C12Q2523/313
摘要： A method for assaying binding between a fluorophore-labeled compound and an unlabeled compound is provided. The method includes detecting a quenching effect on fluorescence emitted by the fluorophore-labeled compound resulting from binding. The binding is specific and other than nucleobase to nucleobase. The method is conducted without separating complexes of the fluorophore-labeled compound and the unlabeled compound from the fluorophore-labeled compound prior to quenching effect detecting, and without providing a signal quenching agent to quench fluorescent light. Preferably, the fluorophore-labeled compound is a nucleic acid and the unlabeled compound is a protein. The method can be used for a variety of applications, including screening for drug candidates having optimum binding properties, and quantifying the binding affinity of DNA binding proteins for nucleic acids.
摘要翻译： 提供了用于测定荧光团标记的化合物和未标记的化合物之间的结合的方法。 该方法包括检测由结合产生的荧光团标记化合物发射的荧光的淬灭效应。 结合是特异性的，而不是核碱基到核碱基。 在淬灭效应检测之前，不从荧光团标记的化合物上分离荧光团标记的化合物和未标记的化合物的复合物，并且不提供信号猝灭剂来淬灭荧光。 优选地，荧光团标记的化合物是核酸，未标记的化合物是蛋白质。 该方法可用于各种应用，包括筛选具有最佳结合特性的候选药物，并定量DNA结合蛋白对核酸的结合亲和力。
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公开(公告)号：US07052844B2
公开(公告)日：2006-05-30
申请号：US10269229
申请日：2002-10-11
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6816 , C12Q1/6827 , C12Q1/6839 , C12Q2563/173 , C12Q2563/107 , C12Q2537/119 , C12Q2527/125 , C12Q2563/137
摘要： A purification method includes bonding a probe to a target in a sample to form a complex, and separating the sample from the complex to separate in a sequence specific manner the target from the sample. The complex, which is immobilized on a support, is a duplex, triplex or quadruplex formed by Watson-Crick complementary base interaction or by homologous base interaction, provided that when the complex is a duplex and the heteropolymeric probe sequence is antiparallel to the heteropolymeric target sequence, the heteropolymeric probe sequence is bonded to the heteropolymeric target sequence by homologous base interaction, and provided that when the complex is a triplex, the complex is free of recombination proteins. A kit for performing the method includes the support and the probe.
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公开(公告)号：US06900300B1
公开(公告)日：2005-05-31
申请号：US09664827
申请日：2000-09-19
申请人： Glen H. Erikson , Jasmine I. Daksis
发明人： Glen H. Erikson , Jasmine I. Daksis
IPC分类号： G01N33/483 , C12N15/09 , C12Q1/68 , G01N33/53 , G01N33/566 , G01N33/58 , C07H21/02 , C07H21/04
CPC分类号： C12Q1/6839 , C12Q2527/125 , C12Q2563/173
摘要： A multiplex structure, such as a nucleic acid quadruplex, includes: a first strand containing a first sequence of nucleobases; a second strand containing a second sequence of nucleobases, wherein the second strand is associated with the first strand by Watson-Crick bonding; a third strand containing a third sequence of nucleobases; and a fourth strand containing a fourth sequence of nucleobases, wherein the fourth strand is associated with the second strand and the third strand by Watson-Crick bonding. Formation of the multiplex structure is promoted by monovalent cations (e.g., sodium and potassium), divalent cations, multivalent cations, intercalating agents and/or molecules known to bind within the minor grooves of nucleic acids. The multiplex structure and the process of forming it have diagnostic, therapeutic, prophylactic and nanoengineering applications.
摘要翻译： 多重结构，例如核酸四链体，包括：含有第一序列核碱基的第一链; 含有第二序列核碱基的第二链，其中所述第二链通过Watson-Crick结合与所述第一链相关; 含有第三序列核碱基的第三链; 和包含第四序列核碱基的第四链，其中所述第四链与所述第二链和所述第三链通过Watson-Crick键结合。 通过一价阳离子（例如钠和钾），二价阳离子，多价阳离子，嵌入剂和/或已知结合在核酸的小槽内的分子来促进多重结构的形成。 多重结构及其形成过程具有诊断，治疗，预防和纳米工程应用。
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6.发明授权Homogenous assay of duplex of triplex hybridization by means of multiple measurements under varied conditions 有权通过在不同条件下的多次测量，三重杂交的双链体的均一测定公开(公告)号：US06265170B1
公开(公告)日：2001-07-24
申请号：US09490273
申请日：2000-01-24
IPC分类号： C12Q168
CPC分类号： C12Q1/6816 , C12Q2563/113 , Y10T436/143333 , C12Q2563/107 , C12Q2523/307 , C12Q2525/107
摘要： The invention provides homogeneous assay methods for nucleic acid hybridization, detection and evaluation. The assay includes obtaining signals from a test sample both before and during the application of a voltage to the test sample and correlating the signals, each of which is indicative of the binding affinity of the probe and the target to each other. The assay enables determining an extent of matching between the probe and the target, as the voltage can be calibrated so as to destabilize significantly any hybridization except perfectly complementary hybridization. The signals whose magnitude is correlated with binding affinity can be electrical conductance and/or fluorescent intensity. Preferably, both signal pairs are measured and compared so as to enhance the reliability of the assay. The assay can detect specific hybridization between single-stranded probes and non-denatured double-stranded targets to form triplexes, thus obviating the need to denature the targets. The assay methods can also be applied to duplex hybridization complexes.
摘要翻译： 本发明提供用于核酸杂交，检测和评价的均一测定方法。 该测定包括在测试样品施加电压之前和期间从测试样品获得信号，并将信号相关联，每个信号指示探针和靶标彼此的结合亲合力。 该测定能够确定探针与靶标之间的匹配程度，因为电压可被校准，以便显着地使除了完全互补杂交之外的任何杂交不稳定。 幅度与结合亲和力相关的信号可以是电导和/或荧光强度。 优选地，测量和比较两个信号对，以增强测定的可靠性。 该测定可以检测单链探针和非变性双链靶标之间的特异性杂交以形成三链体，从而避免了使靶变性的需要。 测定方法也可应用于双链杂交复合物。
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公开(公告)号：US06927027B2
公开(公告)日：2005-08-09
申请号：US09885731
申请日：2001-06-20
CPC分类号： C12Q1/6827 , C12Q1/6816 , C12Q1/6839 , C12Q2563/173 , C12Q2563/107 , C12Q2527/125 , C12Q2563/137
摘要： Heteropolymeric triplexes and quadruplexes and methods for making them; the use of accelerator agents such as cations to create them; the use of fluorescent intercalators and fluorescent probe-bound non-intercalators to detect them.
摘要翻译： 异构复合物和四极体及其制备方法; 使用促进剂如阳离子来形成它们; 使用荧光嵌入剂和荧光探针结合的非嵌入剂来检测它们。
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公开(公告)号：US06911536B1
公开(公告)日：2005-06-28
申请号：US09713177
申请日：2000-11-15
申请人： Glen H. Erikson , Jasmine I. Daksis
发明人： Glen H. Erikson , Jasmine I. Daksis
CPC分类号： C12Q1/6839 , C12Q1/6816 , C12Q1/6827 , C12Q2563/173 , C12Q2563/107 , C12Q2527/125 , C12Q2563/137
摘要： An assay includes catalytic hybridization of targets and cleavable probes to form triplexes and quadruplexes based on Watson-Crick bonding rules. The probes contain scissile linkages that are cleaved by enzymes when hybridized to a target, yielding detectable probe fragments free of the target. The target is recycled to help catalyze the cleavage of additional intact probes to form additional detectable probe fragments, thus amplifying the signal.
摘要翻译： 测定包括靶基因和可切割探针的催化杂交，以形成基于Watson-Crick粘合规则的三联体和四链体。 探针包含与靶标杂交时被酶切割的剪切连接，产生无目标物的可检测的探针片段。 该靶被再循环以帮助催化另外的完整探针的切割以形成另外的可检测的探针片段，从而扩增信号。
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9.发明授权Parallel or antiparallel, homologous or complementary binding of nucleic acids or analogues thereof to form duplex, triplex or quadruplex complexes 有权核酸或其类似物的平行或反向平行，同源或互补结合形成双链体，三链体或四链体复合物公开(公告)号：US07309569B2
公开(公告)日：2007-12-18
申请号：US10293148
申请日：2002-11-12
CPC分类号： C12Q1/6839 , C12Q1/6816 , C12Q1/6827 , C12Q2525/113 , C12Q2525/101 , C12Q2563/173 , C12Q2563/107 , C12Q2537/119 , C12Q2527/125 , C12Q2563/137
摘要： A complex includes: (1) a probe containing a heteropolymeric probe sequence of nucleic acids or nucleic acid analogues; and (2) a target containing a heteropolymeric target sequence of nucleic acids or nucleic acid analogues, wherein: (a) at least one of the probe and the target is purified or synthetic; and (b) the heteropolymeric probe sequence is bonded to the heteropolymeric target sequence by Watson-Crick complementary base interaction or by homologous base interaction, provided that when the complex is a duplex and the heteropolymeric probe sequence is antiparallel to the heteropolymeric target sequence, the heteropolymeric probe sequence is bonded to the heteropolymeric target sequence by homologous base interaction, and provided that when the complex is a triplex, the complex is free of recombination proteins. A method for assaying a target includes detecting formation of the complex.
摘要翻译： 复合物包括：（1）含有核酸或核酸类似物的异源聚合物探针序列的探针; 和（2）含有核酸或核酸类似物的异源聚合物靶序列的靶，其中：（a）所述探针和靶中的至少一个被纯化或合成; 并且（b）异源聚合物探针序列通过Watson-Crick互补碱基相互作用或通过同源碱基相互作用结合到异源聚合物靶序列，条件是当复合物是双链体并且异源聚合物探针序列与异源聚合物靶序列反平行时， 异源聚合物探针序列通过同源碱基相互作用结合到异源聚合物靶序列，并且条件是当复合物是三链体时，复合物不含重组蛋白质。 用于测定靶的方法包括检测复合物的形成。
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10.发明授权Homogeneous assay of biopolymer binding by means of multiple measurements under varied conditions 失效通过在不同条件下进行多次测量，生物聚合物结合的均相测定公开(公告)号：US07220541B2
公开(公告)日：2007-05-22
申请号：US09911047
申请日：2001-07-23
CPC分类号： C12Q1/6825 , C12Q1/6816 , C12Q2563/113 , C12Q2563/107 , C12Q2523/307 , C12Q2525/107
摘要： A method for homogeneously assaying biopolymer bonding includes obtaining signals from a test sample before, during and/or after the application of stimulus to the test sample and correlating the signals. The signals, whose magnitude correlate with binding affinity, can be, for example, electrical conductance and/or fluorescent intensity. The stimulus can be, for example, electric voltage and/or laser radiation. Preferably, different types of signals are measured and compared so as to enhance the reliability of the assay.
摘要翻译： 用于均匀测定生物聚合物键合的方法包括在向测试样品施加刺激之前，期间和/或之后从测试样品获得信号并使信号相关。 与结合亲和力相关的幅度的信号可以是例如电导和/或荧光强度。 刺激可以是例如电压和/或激光辐射。 优选地，测量和比较不同类型的信号，以增强测定的可靠性。
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