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1. 发明授权

US08846314B2 Isotachophoretic focusing of nucleic acids 有权
标题翻译：核酸的同步聚焦聚焦
公开(公告)号：US08846314B2
公开(公告)日：2014-09-30
申请号：US12716142
申请日：2010-03-02
申请人： Robert D. Chambers , Juan G. Santiago , Alexandre Persat , Reto B. Schoch , Mostafa Ronaghi
发明人： Robert D. Chambers , Juan G. Santiago , Alexandre Persat , Reto B. Schoch , Mostafa Ronaghi
IPC分类号： C12Q1/68 , B01D57/02 , C07H21/04 , C12N15/10
CPC分类号： G01N27/447 , B01D57/02 , C12N15/101
摘要： A method and system are presented for fast and efficient isolation, purification and quantitation of nucleic acids from complex biological samples using isotachophoresis in microchannels. In an embodiment, a sieving medium may be used to enhance selectivity. In another embodiment, PCR-friendly chemistries are used to purify nucleic acids from complex biological samples and yield nucleic acids ready for further analysis including for PCR. In another embodiment, small RNAs from biological samples are extracted, isolated, preconcentrated and quantitated using on-chip ITP with a high efficiency sieving medium. The invention enables fast concentration and separation (takes 10s to 100s of seconds) of nucleic acids with high selectivity and using lower volumes of reagents (order of 10s of μL to focus less than 1 pg/μL of nucleic acid).
摘要翻译：提出了一种方法和系统，用于在微通道中使用等速电泳，从复杂生物样品中快速有效地分离，纯化和定量核酸。在一个实施方案中，可以使用筛分介质来增强选择性。在另一个实施方案中，使用PCR友好的化学物质从复杂的生物样品中纯化核酸，并产生准备用于进一步分析的核酸，包括用于PCR。在另一个实施方案中，使用具有高效筛选培养基的片上ITP，提取，分离，预浓缩和定量来自生物样品的小RNA。本发明使得能够以高选择性快速浓缩和分离（需要10秒至100秒）的核酸，并且使用较低体积的试剂（10μL的浓度以聚焦小于1pg /μL的核酸）。

2. 发明授权

US08524061B2 On-chip hybridization coupled with ITP based purification for fast sequence specific identification 有权
标题翻译：片上杂交与基于ITP的纯化相结合，用于快速序列特异性鉴定
公开(公告)号：US08524061B2
公开(公告)日：2013-09-03
申请号：US13373773
申请日：2011-11-29
申请人： Paul J. Utz , Juan G. Santiago , Michael G. Kattah , Alexandre Persat
发明人： Paul J. Utz , Juan G. Santiago , Michael G. Kattah , Alexandre Persat
IPC分类号： G01N27/447 , G01N27/26
CPC分类号： G01N27/44726 , C07K1/26 , C12Q1/68 , C12Q1/6816 , G01N33/6803 , C12Q2525/207 , C12Q2525/301 , C12Q2565/125 , C12Q2565/629
摘要： Isotachophoresis (ITP) can be employed to simultaneously focus the target and ligand of an assay into the same ITP focus zone. The target and ligand can bind to each other in the ITP focus zone, and then the resulting bound complex can be detected (e.g., by fluorescence). The sensitivity of this approach can be greatly increased by the enhanced concentration of both target and ligand that ITP provides in the focus zone. Since ITP can be performed quickly, the resulting assay is both rapid and sensitive. Markers of bacterial urinary tract infections have been experimentally detected at clinically relevant concentrations with this approach. MicroRNA sequences have also been profiled with this approach, which is clinically relevant because MicroRNA is expected to provide useful markers for disease. In one experiment, miR-122 in human kidney and liver was detected and quantified.
摘要翻译：可以使用等速电泳（ITP）来同时将测定的靶和配体聚焦到相同的ITP聚焦区域中。靶和配体可以在ITP聚焦区域中彼此结合，然后可以检测所得到的结合复合物（例如通过荧光）。通过ITP在对焦区域提供的靶和配体的增强浓度，可以大大提高该方法的灵敏度。由于ITP可以快速进行，因此得到的测定既快速又敏感。通过这种方法已经在临床相关浓度上实验检测到细菌性尿路感染的标记。 MicroRNA序列也已经用这种方法进行了描述，这是临床上相关的，因为预期MicroRNA可以为疾病提供有用的标记。在一个实验中，检测并定量人肾脏和肝脏中的miR-122。

3. 发明申请

US20120160689A1 On-chip hybridization coupled with ITP based purification for fast sequence specific identification 有权
标题翻译：片上杂交与基于ITP的纯化相结合，用于快速序列特异性鉴定
公开(公告)号：US20120160689A1
公开(公告)日：2012-06-28
申请号：US13373773
申请日：2011-11-29
申请人： Paul J. Utz , Juan G. Santiago , Michael G. Kattah , Alexandre Persat
发明人： Paul J. Utz , Juan G. Santiago , Michael G. Kattah , Alexandre Persat
IPC分类号： G01N27/447
CPC分类号： G01N27/44726 , C07K1/26 , C12Q1/68 , C12Q1/6816 , G01N33/6803 , C12Q2525/207 , C12Q2525/301 , C12Q2565/125 , C12Q2565/629
摘要： Isotachophoresis (ITP) can be employed to simultaneously focus the target and ligand of an assay into the same ITP focus zone. The target and ligand can bind to each other in the ITP focus zone, and then the resulting bound complex can be detected (e.g., by fluorescence). The sensitivity of this approach can be greatly increased by the enhanced concentration of both target and ligand that ITP provides in the focus zone. Since ITP can be performed quickly, the resulting assay is both rapid and sensitive. Markers of bacterial urinary tract infections have been experimentally detected at clinically relevant concentrations with this approach. MicroRNA sequences have also been profiled with this approach, which is clinically relevant because MicroRNA is expected to provide useful markers for disease. In one experiment, miR-122 in human kidney and liver was detected and quantified.
摘要翻译：可以使用等速电泳（ITP）来同时将测定的靶和配体聚焦到相同的ITP聚焦区域中。靶和配体可以在ITP聚焦区域中彼此结合，然后可以检测所得到的结合复合物（例如通过荧光）。通过ITP在对焦区域提供的靶和配体的增强浓度，可以大大提高该方法的灵敏度。由于ITP可以快速进行，因此得到的测定既快速又敏感。通过这种方法已经在临床相关浓度上实验检测到细菌性尿路感染的标记。 MicroRNA序列也已经用这种方法进行了描述，这是临床上相关的，因为预期MicroRNA可以为疾病提供有用的标记。在一个实验中，检测并定量人肾脏和肝脏中的miR-122。

4. 发明申请

US20120228141A1 LIQUID AND GEL ELECTRODES FOR TRANSVERSE FREE FLOW ELECTROPHORESIS 有权
标题翻译：液体和凝胶电极用于横向自由流动电泳
公开(公告)号：US20120228141A1
公开(公告)日：2012-09-13
申请号：US13413161
申请日：2012-03-06
申请人： Byoungsok Jung , Klint A. Rose , Maxim Shusteff , Alexandre Persat , Juan Santiago
发明人： Byoungsok Jung , Klint A. Rose , Maxim Shusteff , Alexandre Persat , Juan Santiago
IPC分类号： C25B9/00 , B01D43/00
CPC分类号： B01D43/00 , B01D57/02 , G01N27/44769
摘要： The present invention provides a mechanism for separating or isolating charged particles under the influence of an electric field without metal electrodes being in direct contact with the sample solution. The metal electrodes normally in contact with the sample are replaced with high conductivity fluid electrodes situated parallel and adjacent to the sample. When the fluid electrodes transmit the electric field across the sample, particles within the sample migrate according to their electrophoretic mobility.
摘要翻译：本发明提供了一种用于在没有金属电极与样品溶液直接接触的电场的影响下分离或分离带电粒子的机理。通常与样品接触的金属电极被替换为与样品平行且相邻的高导电性流体电极。当流体电极跨越样品传播电场时，样品内的颗粒根据其电泳迁移率而迁移。

5. 发明申请

US20100224494A1 Isotachophoretic Focusing of Nucleic Acids 有权
标题翻译：核酸的同位素聚焦
公开(公告)号：US20100224494A1
公开(公告)日：2010-09-09
申请号：US12716142
申请日：2010-03-02
申请人： Robert D. Chambers , Juan G. Santiago , Alexandre Persat , Reto B. Schoch , Mostafa Ronaghi
发明人： Robert D. Chambers , Juan G. Santiago , Alexandre Persat , Reto B. Schoch , Mostafa Ronaghi
IPC分类号： B01D57/02
CPC分类号： G01N27/447 , B01D57/02 , C12N15/101
摘要： A method and system are presented for fast and efficient isolation, purification and quantitation of nucleic acids from complex biological samples using isotachophoresis in microchannels. In an embodiment, a sieving medium may be used to enhance selectivity. In another embodiment, PCR-friendly chemistries are used to purify nucleic acids from complex biological samples and yield nucleic acids ready for further analysis including for PCR. In another embodiment, small RNAs from biological samples are extracted, isolated, preconcentrated and quantitated using on-chip ITP with a high efficiency sieving medium. The invention enables fast concentration and separation (takes 10s to 100s of seconds) of nucleic acids with high selectivity and using lower volumes of reagents (order of 10s of μL to focus less than 1 pg/μL of nucleic acid).
摘要翻译：提出了一种方法和系统，用于在微通道中使用等速电泳，从复杂生物样品中快速有效地分离，纯化和定量核酸。在一个实施方案中，可以使用筛分介质来增强选择性。在另一个实施方案中，使用PCR友好的化学物质从复杂的生物样品中纯化核酸，并产生准备用于进一步分析的核酸，包括用于PCR。在另一个实施方案中，使用具有高效筛选培养基的片上ITP，提取，分离，预浓缩和定量来自生物样品的小RNA。本发明使得能够以高选择性快速浓缩和分离（需要10秒至100秒）的核酸，并且使用较低体积的试剂（10μL的浓度以聚焦小于1pg /μL的核酸）。

6. 发明申请

US20190271661A1 Enhanced Isotachophoresis Assays Using Additives with Spatial Gradients 审中-公开
公开(公告)号：US20190271661A1
公开(公告)日：2019-09-05
申请号：US16193535
申请日：2018-11-16
申请人： Juan G. Santiago , Alexandre Persat , Giancarlo Garcia , Charbel Eid
发明人： Juan G. Santiago , Alexandre Persat , Giancarlo Garcia , Charbel Eid
IPC分类号： G01N27/447
摘要： Techniques for enhanced isotachophoresis assays using additives with spatial gradients include forming a concentration gradient of an additive along a channel from an input port to an output port. The channel is used for isotachophoresis with ions of a leading electrolyte having a first mobility greater than a mobility of an analyte, and ions of a trailing electrolyte having a second mobility less than the mobility of the analyte. The additive is different from both the leading electrolyte and the trailing electrolyte; and the additive has a third mobility that assures the analyte will encounter the additive. The method further comprises introducing a mixture of the trailing electrolyte and a sample including the analyte. The method further comprises applying an electric field to the channel; and, measuring the analyte.

7. 发明授权

US08999129B2 Liquid and gel electrodes for transverse free flow electrophoresis 有权
标题翻译：用于横向自由电泳的液体和凝胶电极
公开(公告)号：US08999129B2
公开(公告)日：2015-04-07
申请号：US13413161
申请日：2012-03-06
申请人： Byoungsok Jung , Klint A. Rose , Maxim Shusteff , Alexandre Persat , Juan Santiago
发明人： Byoungsok Jung , Klint A. Rose , Maxim Shusteff , Alexandre Persat , Juan Santiago
IPC分类号： G01N27/447 , B01D57/02 , B01D43/00
CPC分类号： B01D43/00 , B01D57/02 , G01N27/44769
摘要： The present invention provides a mechanism for separating or isolating charged particles under the influence of an electric field without metal electrodes being in direct contact with the sample solution. The metal electrodes normally in contact with the sample are replaced with high conductivity fluid electrodes situated parallel and adjacent to the sample. When the fluid electrodes transmit the electric field across the sample, particles within the sample migrate according to their electrophoretic mobility.
摘要翻译：本发明提供了一种用于在没有金属电极与样品溶液直接接触的电场的影响下分离或分离带电粒子的机理。通常与样品接触的金属电极被替换为与样品平行且相邻的高导电性流体电极。当流体电极跨越样品传播电场时，样品内的颗粒根据其电泳迁移率而迁移。

8. 发明申请

US20100084271A1 Control of chemical reactions using isotachophoresis 有权
标题翻译：使用等速电泳控制化学反应
公开(公告)号：US20100084271A1
公开(公告)日：2010-04-08
申请号：US12587537
申请日：2009-10-07
申请人： Juan G. Santiago , Alexandre Persat
发明人： Juan G. Santiago , Alexandre Persat
IPC分类号： G01N27/26
CPC分类号： G01N27/44765 , B01D57/02 , C12M1/42 , C12P19/34 , C12Q1/686 , G01N27/447 , G01N27/44721 , G01N27/44791 , G01N27/44795 , C12Q2565/629 , C12Q2527/125
摘要： Isotachophoresis (ITP) is exploited to control various aspects of chemical reactions. In a first aspect, at least one of the reactants of a chemical reaction is confined to an ITP zone, but the resulting product of the chemical reaction is separated from this ITP zone by the ITP process. In a second aspect, one or more reactants of a chemical reaction are confined to an ITP zone, and one or more other reactants of the chemical reaction are not confined to this ITP zone. In a third aspect, ITP is employed to confine at least one reactant of a chemical reaction to an ITP zone, and at least one reactant of the chemical reaction is delivered to the ITP zone in two or more discrete doses. These aspects are especially relevant to performing polymerase chain reactions using chemical denaturants as opposed to thermal cycling.
摘要翻译：利用等速电泳（ITP）来控制化学反应的各个方面。在第一方面，化学反应的至少一种反应物被限制在ITP区域，但化学反应的所得产物通过ITP方法与该ITP区域分离。在第二方面，一种或多种化学反应的反应物被限制在ITP区域，化学反应的一种或多种其它反应物不限于该ITP区域。在第三方面，ITP用于将至少一种化学反应的反应物限制于ITP区，并且化学反应的至少一种反应物以两个或更多个离散剂量递送至ITP区。这些方面与使用化学变性剂而不是热循环进行聚合酶链反应特别相关。

9. 发明授权

US10132775B2 Enhanced isotachophoresis assays using additives with spatial gradients 有权
公开(公告)号：US10132775B2
公开(公告)日：2018-11-20
申请号：US14828487
申请日：2015-08-17
申请人： Juan G. Santiago , Alexandre Persat , Giancarlo Garcia , Charbel Eid
发明人： Juan G. Santiago , Alexandre Persat , Giancarlo Garcia , Charbel Eid
IPC分类号： G01N27/447
摘要： Techniques for enhanced isotachophoresis assays using additives with spatial gradients include forming a concentration gradient of an additive along a channel from an input port to an output port. The channel is used for isotachophoresis with ions of a leading electrolyte having a first mobility greater than a mobility of an analyte, and ions of a trailing electrolyte having a second mobility less than the mobility of the analyte. The additive is different from both the leading electrolyte and the trailing electrolyte; and the additive has a third mobility that assures the analyte will encounter the additive. The method further comprises introducing a mixture of the trailing electrolyte and a sample including the analyte. The method further comprises applying an electric field to the channel; and, measuring the analyte.

10. 发明申请

US20160153934A1 Enhanced Isotachophoresis Assays Using Additives with Spatial Gradients 审中-公开
标题翻译：使用添加剂与空间梯度增强等速电泳分析
公开(公告)号：US20160153934A1
公开(公告)日：2016-06-02
申请号：US14828487
申请日：2015-08-17
申请人： Juan G. Santiago , Alexandre Persat , Giancarlo Garcia , Charbel Eid
发明人： Juan G. Santiago , Alexandre Persat , Giancarlo Garcia , Charbel Eid
IPC分类号： G01N27/447
CPC分类号： G01N27/44756 , G01N27/44721 , G01N27/4473 , G01N27/44795
摘要： Techniques for enhanced isotachophoresis assays using additives with spatial gradients include forming a concentration gradient of an additive along a channel from an input port to an output port. The channel is used for isotachophoresis with ions of a leading electrolyte having a first mobility greater than a mobility of an analyte, and ions of a trailing electrolyte having a second mobility less than the mobility of the analyte. The additive is different from both the leading electrolyte and the trailing electrolyte; and the additive has a third mobility that assures the analyte will encounter the additive. The method further comprises introducing a mixture of the trailing electrolyte and a sample including the analyte. The method further comprises applying an electric field to the channel; and, measuring the analyte.
摘要翻译：使用具有空间梯度的添加剂进行增强等速电泳测定的技术包括沿着通道从输入端口到输出端口形成添加剂的浓度梯度。该通道用于具有第一迁移率大于分析物的迁移率的前导电解质的离子和具有小于分析物的迁移率的第二迁移率的尾电解质的离子的等速电泳。添加剂不同于主要电解质和尾电解质; 并且添加剂具有确保分析物将遇到添加剂的第三迁移率。该方法还包括引入拖尾电解质和包括分析物的样品的混合物。该方法还包括向信道施加电场; 并测量分析物。

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