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1. 发明申请

US20130323729A1 Proximity Ligation Technology for Western Blot Applications 审中-公开
标题翻译：西印迹应用的接近连接技术
公开(公告)号：US20130323729A1
公开(公告)日：2013-12-05
申请号：US13882228
申请日：2011-10-27
申请人： Ulf Landegren , Åsa Hagner-McWhirter , Daniel Ivansson
发明人： Ulf Landegren , Åsa Hagner-McWhirter , Daniel Ivansson
IPC分类号： G01N33/543
CPC分类号： G01N33/5436 , C12Q1/6804 , C12Q1/682 , G01N33/542 , G01N33/561 , G01N33/6803 , G01N2458/10 , C12Q2531/125 , C12Q2561/125 , C12Q2521/501 , C12Q2563/107 , C12Q2563/125
摘要： The invention provides a method for detecting a biomolecular feature (a protein, protein complex, or modified protein such as a phosphorylated protein) by a modified Western blot type of assay, which method either electrophoretic gel separation followed by transfer, or direct spotting of a sample containing the biomolecular feature onto a membrane; providing a proximity probe pair, each probe comprising a binding moiety with affinity for a different binding site on the bio molecular feature and a reactive oligonucleotide, coupled thereto; binding the proximity probes to their respective binding sites on the biomolecular feature through the binding moiety, adding a splint oligonucleotide and a backbone oligonucleotide which are complementary to the reactive oligonucleotide pair, and allowing hybridization among them; ligating the hybridized DNA oligonucleotides to create a circularized DNA molecule when both probes bind sufficiently close to each other on the bio molecular feature, amplifying the circularized DNA by isothermal amplification; and detecting the presence and quantity of the bio molecular feature using a detection oligonucleotide complementary to the amplification product. Also provided are methods for multiplexed detection of more than one bio molecular feature, as well as kits for performing the assays.
摘要翻译：本发明提供了通过修饰的Western印迹分析法检测生物分子特征（蛋白质，蛋白质复合物或修饰蛋白质，例如磷酸化蛋白质）的方法，该方法是电泳凝胶分离，然后转移，或直接点样含有生物分子特征的样品到膜上; 提供接近探针对，每个探针包含对生物分子特征上的不同结合位点具有亲和力的结合部分和与其偶联的反应性寡核苷酸; 通过结合部分将接近探针结合到其生物分子特征上的各自的结合位点，加入与反应性寡核苷酸对互补的夹板寡核苷酸和主链寡核苷酸，并允许它们之间的杂交; 当两个探针在生物分子特征上彼此足够接近时，连接杂交的DNA寡核苷酸以产生环化的DNA分子，通过等温扩增扩增环化的DNA; 并使用与扩增产物互补的检测寡核苷酸检测生物分子特征的存在和数量。还提供了用于多个检测多于一个生物分子特征的方法以及用于进行测定的试剂盒。

2. 发明授权

US08053188B2 Nucleic acid enrichment 有权
标题翻译：核酸浓缩
公开(公告)号：US08053188B2
公开(公告)日：2011-11-08
申请号：US10495895
申请日：2002-11-20
申请人： Mats Gullberg , Ulf Landegren
发明人： Mats Gullberg , Ulf Landegren
IPC分类号： C12Q1/68 , C07H21/04
CPC分类号： C12Q1/6827 , C12Q1/683 , C12Q1/6858 , C12Q2561/125 , C12Q2525/137 , C12Q2521/319 , C12Q2535/131 , C12Q2521/313 , C12Q2531/125
摘要： This invention relates to methods, reagents and kits for enriching nucleic acid sequences. More particularly, the present invention relates to methods, reagents and kits for sample preparation including sample modification, sample enrichment and amplification.
摘要翻译：本发明涉及用于富集核酸序列的方法，试剂和试剂盒。更具体地，本发明涉及用于样品制备的方法，试剂和试剂盒，包括样品修饰，样品富集和扩增。

3. 发明申请

US20130171652A1 Blocking Reagent and Methods for the Use Thereof 有权
标题翻译：阻塞试剂及其使用方法
公开(公告)号：US20130171652A1
公开(公告)日：2013-07-04
申请号：US13809964
申请日：2011-07-13
申请人： Simon Fredriksson , Bonnie Tran
发明人： Simon Fredriksson , Bonnie Tran
IPC分类号： C12Q1/68
CPC分类号： C12Q1/686 , C12Q1/6804 , G01N33/542 , G01N33/54393 , C12Q2537/163 , C12Q2533/107
摘要： The present invention relates to the use of a conjugate of a non-analyte-specific binding protein coupled to a nucleic acid as a blocking reagent in a probe-based detection assay, which uses a probe comprising a proteinaceous analyte-binding partner coupled to a nucleic acid domain to detect an analyte in a sample.
摘要翻译：本发明涉及在基于探针的检测测定中使用与核酸偶联的非分析物特异性结合蛋白作为封闭试剂的缀合物，其使用包含与蛋白质分析物结合配对物偶联的探针核酸结构域以检测样品中的分析物。

4. 发明授权

US07306904B2 Methods and kits for proximity probing 有权
标题翻译：邻近探测方法和试剂盒
公开(公告)号：US07306904B2
公开(公告)日：2007-12-11
申请号：US09785657
申请日：2001-02-20
申请人： Ulf Landegren , Simon Fredriksson
发明人： Ulf Landegren , Simon Fredriksson
IPC分类号： C12Q1/68 , C07H21/04 , C07K14/00
CPC分类号： G01N33/58 , C12Q1/68 , C12Q1/6804 , G01N33/542 , C12Q2563/179
摘要： The present invention relates to sensitive, rapid and convenient assays for detection and/or quantification of one or several analyte(s) in solution using so called proximity probes. The proximity probes comprise a binding moiety and a nucleic acid. The nucleic acid from one proximity probe is only capable of interaction with the nucleic acid from the other proximity probe when these are in close proximity, i.e. have bound to the analytes for which they are specific. The present invention relates to methods and kits for proximity probing and are performed in solution without the need of a solid phase.
摘要翻译：本发明涉及使用所谓的邻近探针在溶液中检测和/或定量一种或几种分析物的灵敏，快速和方便的测定。邻近探针包含结合部分和核酸。来自一个邻近探针的核酸仅当它们处于非常接近的位置时才能与来自其它接近探针的核酸相互作用，即与其特异性的分析物结合。本发明涉及用于邻近探测的方法和试剂盒，并且在溶液中进行而不需要固相。

5. 发明申请

US20140170654A1 Unfolding Proximity Probes and Methods for the Use Thereof 有权
标题翻译：展开接近探头及其使用方法
公开(公告)号：US20140170654A1
公开(公告)日：2014-06-19
申请号：US14116706
申请日：2012-05-11
申请人： Ulf Landegren , Rachel Yuan Nong , Ola Söderberg , Irene Weibrecht
发明人： Ulf Landegren , Rachel Yuan Nong , Ola Söderberg , Irene Weibrecht
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6876 , C12Q1/6816 , C12Q1/6841 , G01N33/542 , C12Q2521/501 , C12Q2521/531 , C12Q2525/301 , C12Q2525/307 , C12Q2531/125 , C12Q2537/162
摘要： The present invention relates to a proximity-probe based detection assay for detecting an analyte in a sample and in particular to a method that comprises the use of at least one set of at least first and second proximity probes, which probes each comprise an analyte-binding domain and a nucleic acid domain and can simultaneously bind to the analyte directly or indirectly, wherein the nucleic acid domain of at least one of said proximity probes comprises a hairpin structure that can be unfolded by cleavage of the nucleic acid domain to generate at least one ligatable free end or region of complementarity to another nucleic acid molecule in said sample, wherein when the probes bind to said analyte unfolding said hairpin structure allows the nucleic acid domains of said at least first and second proximity probes to interact directly or indirectly.
摘要翻译：本发明涉及一种用于检测样品中分析物的基于接近探针的检测测定法，特别涉及一种包括使用至少一组至少第一和第二接近探针的方法，该探针各自包含分析物 - 结合结构域和核酸结构域，并且可以直接或间接地同时结合分析物，其中至少一个所述邻近探针的核酸结构域包含发夹结构，所述发夹结构可以通过切割核酸结构域而被解折叠以产生至少一个可连接的自由端或与所述样品中的另一个核酸分子互补的区域，其中当所述探针结合到所述分析物上展开所述发夹结构时，允许所述至少第一和第二接近探针的核酸结构域直接或间接相互作用。

6. 发明授权

US08080393B2 Methods for production of oligonucleotides 有权
标题翻译：生产寡核苷酸的方法
公开(公告)号：US08080393B2
公开(公告)日：2011-12-20
申请号：US11911521
申请日：2006-04-10
申请人： Jorn Erland Koch , Magnus Stougaard , Jakob Schwalbe Lohmann
发明人： Jorn Erland Koch , Magnus Stougaard , Jakob Schwalbe Lohmann
IPC分类号： C12P19/34 , C07H21/02
CPC分类号： C12P19/34 , C12N15/10 , C12Q1/6844 , C12Q2531/125 , C12Q2525/131 , C12Q2537/149 , C12Q2525/301 , C12Q2521/307
摘要： The present invention relates to production of oligonucleotides using rolling circle replication, wherein synthesised multimeric oligonucleotides are reduced to mononucleotides using a nicking cassette. Thus, the invention provides a method for the production of oligonucleotides, enabling efficient amplification of oligonucleotides at lengths up to at least 1000 nucleotides and in high amounts contained within a nicking cassette.
摘要翻译：本发明涉及使用滚环复制制备寡核苷酸，其中合成的多聚寡核苷酸使用切割盒被还原成单核苷酸。因此，本发明提供了一种生产寡核苷酸的方法，能够有效扩增长度至多为1000个核苷酸的寡核苷酸，并以高含量包含在切口盒中。

7. 发明申请

US20110223585A1 ASSAY FOR LOCALIZED DETECTION OF ANALYTES 审中-公开
标题翻译：用于本地化检测分析的测定
公开(公告)号：US20110223585A1
公开(公告)日：2011-09-15
申请号：US13045399
申请日：2011-03-10
申请人： MATS GULLBERG , SIMON FREDRIKSSON
发明人： MATS GULLBERG , SIMON FREDRIKSSON
IPC分类号： C12Q1/70 , C12Q1/68
CPC分类号： C12Q1/682 , C12Q1/6804 , C12Q2565/101 , C12Q2563/125 , C12Q2531/125 , C12Q2521/101 , C12Q2563/179 , C12Q2521/301 , C12Q2521/501
摘要： The present invention relates to a method for detecting an analyte in a sample, said method comprising: (a) contacting said sample with at least one set of at least first and second proximity probes, wherein said probes each comprise an analyte-binding moiety and can simultaneously bind to the analyte, and wherein (i) said first proximity probe comprises a nucleic acid moiety attached at one end to the analyte-binding moiety, wherein a circular or circularizable oligonucleotide is hybridized to said nucleic acid moiety before, during or after said contacting step; and (ii) said second proximity probe comprises an enzyme moiety, attached to the analyte-binding moiety, capable of directly or indirectly enabling rolling circle amplification (RCA) of the circular or, when it is circularized, of the circularizable oligonucleotide hybridized to the nucleic acid moiety of the first proximity probe, wherein said RCA is primed by said nucleic acid moiety of said first proximity probe; (b) if necessary, circularizing said oligonucleotide, to produce a circularized template for RCA; (c) subjecting said circular or circularized template to RCA, wherein if the enzyme moiety of the second proximity probe in step (a)(ii) is a DNA polymerase, this step does not utilize a free DNA polymerase; and (d) detecting a product of said RCA.
摘要翻译：本发明涉及一种用于检测样品中分析物的方法，所述方法包括：（a）使所述样品与至少一组至少第一和第二接近探针接触，其中所述探针各自包含分析物结合部分，可以同时结合分析物，并且其中（i）所述第一邻近探针包含在一端与分析物结合部分连接的核酸部分，其中环状或可环化的寡核苷酸在所述核酸部分之前，期间或之后杂交说接触步骤; 和（ii）所述第二邻近探针包含连接到分析物结合部分的酶部分，能够直接或间接地实现圆环的滚环扩增（RCA），或者当环化时，可环化的寡核苷酸与所述第一邻近探针的核酸部分，其中所述RCA由所述第一邻近探针的所述核酸部分引发; （b）如有必要，使所述寡核苷酸通化，以产生RCA的环化模板; （c）使所述圆形或环化模板经受RCA，其中如果步骤（a）（ii）中第二邻近探针的酶部分是DNA聚合酶，则该步骤不使用游离DNA聚合酶; 和（d）检测所述RCA的乘积。

8. 发明授权

US07883848B2 Regulation analysis by cis reactivity, RACR 有权
标题翻译：通过顺式反应性进行调节分析，RACR
公开(公告)号：US07883848B2
公开(公告)日：2011-02-08
申请号：US11483825
申请日：2006-07-10
申请人： Olof Ericsson
发明人： Olof Ericsson
IPC分类号： C12Q1/68 , C12P19/34 , C07H21/02 , G01N33/566
CPC分类号： C12Q1/6804 , C12N15/1075 , G01N33/53 , G01N33/58 , G01N2458/10
摘要： Methods of detecting affinity interactions between at least two molecules of interest are provided. The method comprises: a. forming a plurality of interactors by coupling each molecule of interest with at least one nucleic acid moiety comprising an identification sequence element and at an association element; b. promoting an association between at least two nucleic acid moieties from different interactors to form a plurality of unique associated oligonucleotides, wherein each nucleic acid moiety may form more than one unique associated oligonucleotide, and wherein each unique associated oligonucleotide comprises at least two identification sequence elements derived from the at least two nucleic acid moieties; c. selecting the plurality of unique associated oligonucleotides; and d. subjecting the selected associated oligonucleotides to an analysis that permits detection of the at least two identification sequence elements. Similar methods directed to detecting functional interactions, libraries of interactors employable in the present methods, and kits comprising those libraries are also provided.
摘要翻译：提供了检测至少两个感兴趣的分子之间的亲和力相互作用的方法。该方法包括：a。通过将每个感兴趣的分子与至少一个包含识别序列元件的核酸部分和缔合元件相耦合来形成多个相互作用体; b。促进来自不同相互作用者的至少两个核酸部分之间的缔合以形成多个独特的相关寡核苷酸，其中每个核酸部分可以形成多于一个唯一相关的寡核苷酸，并且其中每个唯一相关寡核苷酸包含至少两个识别序列元件从所述至少两个核酸部分; C。选择所述多个独特的相关寡核苷酸; 和d。对所选择的相关寡核苷酸进行允许检测所述至少两个识别序列元件的分析。还提供了用于检测功能相互作用的类似方法，在本方法中可使用的相互作用者的文库以及包括这些文库的试剂盒。

9. 发明申请

US20100021890A1 METHOD FOR ANALYTE DETECTION USING PROXIMITY PROBES 有权
标题翻译：使用临近探针进行分析检测的方法
公开(公告)号：US20100021890A1
公开(公告)日：2010-01-28
申请号：US12294031
申请日：2007-03-20
申请人： Edith Schallmeiner
发明人： Edith Schallmeiner
IPC分类号： C12Q1/68 , G01N33/566
CPC分类号： C12Q1/682 , G01N33/542 , C12Q2563/131 , C12Q2537/125 , C12Q2533/107
摘要： The present invention relates to a method for detecting an analyte in a sample, comprising (a) contacting said sample with at least one set of at least first, second and third proximity probes, which probes each comprise an analyte-binding domain and a nucleic acid domain and can simultaneously bind to the analyte, the nucleic acid domain of said third proximity probe being a splint which is capable of hybridising at least to the nucleic acid domains of said first and second proximity probes, wherein when all of the at least three proximity probes bind to said analyte, the nucleic acid domains of said first and second proximity probes are conjugatable by means of an interaction mediated by said hybridised splint of said third proximity probe; (b) conjugating the nucleic acids, of said first and second proximity probes; and (c) detecting said conjugation. Also provided is a kit for use in such a method.
摘要翻译：本发明涉及一种用于检测样品中的分析物的方法，其包括（a）使所述样品与至少一组至少第一，第二和第三接近度探针接触，所述探针各自包含分析物结合结构域和核酸并且可以同时结合分析物，所述第三邻近探针的核酸结构域是能够至少与所述第一和第二接近探针的核酸结构域杂交的夹板，其中当所述至少三个邻近探针结合所述分析物，所述第一和第二接近探针的核酸结构域可通过由所述第三接近探针的所述杂交夹板介导的相互作用而共轭; （b）使所述第一和第二接近探针的核酸缀合; 和（c）检测所述共轭。还提供了用于这种方法的试剂盒。

10. 发明申请

US20160376642A1 Localised RCA-based Amplification Method 审中-公开
标题翻译：基于RCA的局部放大方法
公开(公告)号：US20160376642A1
公开(公告)日：2016-12-29
申请号：US14442701
申请日：2013-11-14
申请人： OLINK AB
发明人： Ulf LANDEGREN , Lei CHEN , Di WU , Yuan NONG , Caroline GALLANT
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6848 , C12Q1/6804 , C12Q1/682 , C12Q2531/125 , C12Q2537/149 , C12Q2533/107 , C12Q2525/131 , C12Q2525/301 , C12Q2525/155
摘要： The present invention provides a method for performing a localised RCA reaction comprising at least two rounds of RCA, wherein the product of a second RCA reaction is attached, and hence localised, to a product of a first RCA reaction, said method comprising: (a) providing a first RCA product; (b) directly or indirectly hybridising to said first RCA product a probe which comprises or provides a primer for a second RCA reaction; and (c) performing a second RCA reaction using said RCA primer of (b) to form a second RCA product, wherein in said reaction: (i) said probe and said primer are not able to prime extension using said first RCA product as template or any such extension is limited to avoid displacement of any probe hybridised to the first RCA product; (ii) the direct or indirect hybridisation of the RCA primer of (b) to the first RCA product is maintained and, by virtue of said hybridisation, the second RCA product is attached to the first RCA product; (iii) a RCA template for said second RCA reaction is comprised in or provided by the probe, or is separately provided. The method finds particular utility in the detection of analytes, wherein the analyte is a nucleic acid or wherein a nucleic acid is used or generated as a marker for the analyte.
摘要翻译：本发明提供了一种执行包含至少两轮RCA的局部RCA反应的方法，其中将第二RCA反应的产物连接并因此定位于第一RCA反应的产物，所述方法包括：（a ）提供第一个RCA产品; （b）与所述第一RCA产物直接或间接杂交包含或提供用于第二RCA反应的引物的探针; （c）使用（b）的所述RCA引物进行第二RCA反应以形成第二RCA产物，其中在所述反应中：（i）所述探针和所述引物不能使用所述第一RCA产物作为模板进行扩展或者任何这样的扩展被限制以避免杂交到第一RCA产品的任何探针的位移; （ii）维持（b）的RCA引物与第一RCA产物的直接或间接杂交，并且由于所述杂交，第二RCA产物连接到第一RCA产物; （iii）用于所述第二RCA反应的RCA模板包含在探针中或由探针提供，或单独提供。该方法在检测分析物中具有特别的用途，其中分析物是核酸，或者其中使用或产生核酸作为分析物的标记物。

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