公开(公告)号：US07871519B2

公开(公告)日：2011-01-18

申请号：US11910057

申请日：2006-03-31

申请人： Christine Kurata ,  Daniel Capaldi ,  Zhiwei Wang ,  Nhuy Luu ,  Hans-joachim Gaus ,  Claus André Frank Rentel

发明人： Christine Kurata ,  Daniel Capaldi ,  Zhiwei Wang ,  Nhuy Luu ,  Hans-joachim Gaus ,  Claus André Frank Rentel

IPC分类号： B01D15/08

CPC分类号： G01N30/7233 ,  G01N30/02 ,  G01N30/34 ,  G01N2030/342 ,  G01N2030/8813 ,  G01N2030/8827 ,  B01D15/366

摘要： Provided is a method comprising introducing a sample comprising a plurality of oligonucleotides into an ion pair high performance liquid chromatography column having a buffered mobile phase and allowing at least a portion of the oligonucleotides to separate; allowing the oligonucleotides to elute from the column; and introducing the oligonucleotides into a mass spectrometer and quantifying at least a portion of the oligonucleotides by mass spectrometry. In the method, at least a portion of the oligonucleotides are co-eluting oligonucleotides that differ in mass by no more than 20%; and the buffered mobile phase causes at least 50 mole percent of co-eluting oligonucleotides to have the same charge when they enter the mass spectrometer.

摘要翻译： 提供了一种方法，包括将包含多个寡核苷酸的样品引入具有缓冲流动相的离子对高效液相色谱柱中，并允许至少一部分寡核苷酸分离; 允许寡核苷酸从柱洗脱; 并将寡核苷酸引入质谱仪中，并通过质谱法定量至少一部分寡核苷酸。 在该方法中，寡核苷酸的至少一部分是质量不超过20％的共洗脱寡核苷酸; 并且缓冲的流动相使至少50摩尔％的共洗脱寡核苷酸在进入质谱仪时具有相同的电荷。

公开(公告)号：US07169298B2

公开(公告)日：2007-01-30

申请号：US09770410

申请日：2001-01-25

申请人： Christian Huber ,  Herbert Oberarcher ,  Andreas Premstaller

发明人： Christian Huber ,  Herbert Oberarcher ,  Andreas Premstaller

IPC分类号： B01D15/08

CPC分类号： B01J20/285 ,  B01D15/366 ,  B01J20/26 ,  B01J20/261 ,  B01J20/28014 ,  B01J20/28042 ,  B01J20/281 ,  B01J20/287 ,  B01J20/3064 ,  B01J20/327 ,  B01J2220/54 ,  B01J2220/82 ,  B01J2220/84 ,  G01N30/02 ,  G01N30/48 ,  G01N2030/525 ,  G01N2030/528

摘要： Methods and devices based on capillary monolithic columns, preferably consisting of an underivatized poly(styrene-divinylbenzene) monolith, for separating a mixture of polynucleotides by ion pair-reverse phase-high performance chromatography (IP-RP-HPLC). In various aspects of the method and device the monolith is characterized by one or more of the following: the monolith is contained within a capillary tube; the monolith is immobilized by covalent attachment at the inner wall of the tube; the tube is devoid of retaining frits; the monolith is characterized by having above 10,000 theoretical plates per meter and preferably above 200,000 theoretical plates per meter; the method uses a mobile phase which is devoid of EDTA; the monolith has a surface morphology that is rugulose or brush-like; the chromatographic surfaces of the monolith are non-porous; the monolith has channels sufficiently large for convective flow of the mobile phase; the monolith is formed from a polymerization mixture including underivatized styrene, a crosslinking agent, and a porogen, wherein the porogen includes tetrahydrofuran. The monolith can be incorporated into a miniaturized chromatography system which can be coupled to a mass spectrometer for on-line separation and mass determination of single- or double-stranded polynucleotides.

摘要翻译： 基于毛细管整体柱的方法和装置，优选由未衍生的聚（苯乙烯 - 二乙烯基苯）整料组成，用于通过离子对 - 反相 - 高性能色谱法（IP-RP-HPLC）分离多核苷酸的混合物。 在该方法和装置的各个方面，整料的特征在于以下一个或多个：整料包含在毛细管内; 通过在管的内壁上共价连接将整料固定化; 管没有保留玻璃料; 该单块的特征在于每米具有超过10,000理论塔板，优选地每平方米超过20万理论塔板; 该方法使用无EDTA的流动相; 整料具有红褐色或刷状的表面形态; 整料的色谱表面是无孔的; 整体结构具有足够大的流动相的对流流动通道; 整料由包含未衍生的苯乙烯，交联剂和致孔剂的聚合混合物形成，其中致孔剂包括四氢呋喃。 整料可以并入小型化的色谱系统中，该系统可与质谱联用，用于单链或双链多核苷酸的在线分离和质量测定。

公开(公告)号：US07135289B2

公开(公告)日：2006-11-14

申请号：US10266906

申请日：2002-10-07

申请人： Paul D. Taylor ,  Liem T. Nguyen

发明人： Paul D. Taylor ,  Liem T. Nguyen

IPC分类号： C12Q1/68 ,  A61K7/075 ,  A01N57/26

CPC分类号： G01N30/34 ,  B01D15/161 ,  B01D15/20 ,  B01D15/325 ,  B01D15/366 ,  B01D15/426 ,  C12Q1/6827 ,  G01N30/02 ,  C12Q2565/137 ,  C12Q2527/125 ,  G01N2030/3007 ,  C12Q2545/113 ,  G01N30/30

摘要： Methods, compositions, and kits for separating heteroduplex and homoduplex DNA molecules in a test mixture by temperature-compression denaturing high performance liquid chromatography (tcDHPLC). The method includes use of nitrogen-containing additives in the mobile phase that allow detection of diverse heteroduplex molecules to be performed at the same pre-selected temperature. An example of a preferred additive is betaine. Standard mixtures of DNA fragments, such as mutation standards containing known heteroduplex and homoduplex molecules, can be used to select the concentration of additive and temperature. Compositions and kits including the mobile phase, mutation standards, PCR primers, separation media, and DNA polymerase are also provided.

公开(公告)号：US06746608B2

公开(公告)日：2004-06-08

申请号：US10317980

申请日：2002-12-12

申请人： Leonard H. Smiley

发明人： Leonard H. Smiley

IPC分类号： B01D1500

CPC分类号： B01J20/28019 ,  B01D15/325 ,  B01D15/34 ,  B01D15/361 ,  B01D15/366 ,  B01D15/3804 ,  B01J20/261 ,  B01J20/28033 ,  B01J20/285 ,  B01J20/286 ,  B01J20/287 ,  B01J20/3268 ,  B01J2220/82 ,  C12N15/1006 ,  C12N15/101

摘要： The invention is directed to fluorinated particles having adsorbent properties for superior performance as the stationary phase for use in chromatographic separations. In particular, the fluorinated surface of such particles of the invention presents unusual and unexpected polarity that is beneficial in performing chromatographic separations such as that used for DNA.

摘要翻译： 本发明涉及具有吸附性能的氟化颗粒，其性能优于用于色谱分离的固定相。 特别地，本发明的这种颗粒的氟化表面具有异常和意想不到的极性，这有利于进行色谱分离，例如用于DNA的色谱分离。

公开(公告)号：US06573307B1

公开(公告)日：2003-06-03

申请号：US09980901

申请日：2002-04-15

申请人： Leonard H. Smiley ,  Christopher Lowe ,  Julie Tucker

发明人： Leonard H. Smiley ,  Christopher Lowe ,  Julie Tucker

IPC分类号： C08J926

CPC分类号： B01D15/325 ,  B01D15/1807 ,  B01D15/366 ,  B01D15/3804 ,  B01D2215/021 ,  B01J20/26 ,  B01J20/286 ,  B01J20/287 ,  C12N15/1006 ,  C12N15/101 ,  G01N2030/042 ,  Y10S435/803

摘要： A process to make particles by anaerobic reaction of a water-insoluble solution of organic compounds comprising (a) a monomer selected from C2-4 alkylene glycol esters of a C3-6 acrylic acid and a divinyl benzene; (b) a polyfluorinated vinyl monomer; (c) a monomer selected from acrylic acid, methacrylic acid and esters thereof; (d) a free radical initiator; and (e) a water-insoluble, organic solvent-soluble porogenic material, the weight ratio of comonomers (a) plus (b) plus (c) to the porogenic material being from 0.5:1 to 2:1. The adsorbent particles produced by the process are useful in carrying out chromatographic separations, or in the production of medical devices.

摘要翻译： 一种通过水不溶性有机化合物溶液的厌氧反应制备颗粒的方法，包括（a）选自C 3-6亚烷基二醇酯和二乙烯基苯的单体; （b）多氟乙烯基单体; （c）选自丙烯酸，甲基丙烯酸及其酯的单体; （d）自由基引发剂; 和（e）水不溶性有机溶剂可溶性致孔材料，共聚单体（a）加（b）加（c）与致孔材料的重量比为0.5:1至2:1。 通过该方法生产的吸附剂颗粒可用于进行色谱分离或用于生产医疗器械。

公开(公告)号：US20020164589A1

公开(公告)日：2002-11-07

申请号：US09756070

申请日：2001-01-06

发明人： Paul D. Taylor

IPC分类号： C12Q001/68 ,  C07H021/04

CPC分类号： B01J41/20 ,  B01D15/366 ,  C12Q1/6827 ,  C12Q2565/137

摘要： The present invention describes a method for separating or partially separating heteroduplex and homoduplex DNA molecules in a mixture. In the method, the mixture is applied to an anion-exchange chromatography medium. The heteroduplex and homoduplex molecules are eluted with a mobile phase containing an eluting salt, including an anion and a cation, a buffer, and preferably including an organic solvent. The eluting is carried out under conditions effective to at least partially denature the heteroduplexes (e.g., thermal or chemical denaturing) resulting in the separation of the heteroduplexes from the homoduplexes. The method has many applications including, but not limited to, detecting mutations and comparative DNA sequencing.

摘要翻译： 本发明描述了在混合物中分离或部分分离异源双链体和同源双链DNA分子的方法。 在该方法中，将混合物施加到阴离子交换色谱介质上。 异源二聚体和同源双链分子用含有洗脱盐的流动相洗脱，包括阴离子和阳离子，缓冲液，优选包括有机溶剂。 洗脱在有效地使异源双链体（例如，热或化学变性）至少部分变性的条件下进行，导致异源双链体与同源双链体分离。 该方法具有许多应用，包括但不限于检测突变和比较DNA测序。

公开(公告)号：US20020088753A1

公开(公告)日：2002-07-11

申请号：US09770410

申请日：2001-01-25

发明人： Christian Huber ,  Herbert Oberacher ,  Andreas Premstaller

IPC分类号： B01D015/08

CPC分类号： B01J20/285 ,  B01D15/366 ,  B01J20/26 ,  B01J20/261 ,  B01J20/28014 ,  B01J20/28042 ,  B01J20/281 ,  B01J20/287 ,  B01J20/3064 ,  B01J20/327 ,  B01J2220/54 ,  B01J2220/82 ,  B01J2220/84 ,  G01N30/02 ,  G01N30/48 ,  G01N2030/525 ,  G01N2030/528

摘要： Methods and devices based on capillary monolithic columns, preferably consisting of an underivatized poly(styrene-divinylbenzene) monolith, for separating a mixture of polynucleotides by ion pair-reverse phase-high performance chromatography (IP-RP-HPLC). In various aspects of the method and device the monolith is characterized by one or more of the following: the monolith is contained within a capillary tube; the monolith is immobilized by covalent attachment at the inner wall of the tube; the tube is devoid of retaining frits; the monolith is characterized by having above 10,000 theoretical plates per meter and preferably above 200,000 theoretical plates per meter; the method uses a mobile phase which is devoid of EDTA; the monolith has a surface morphology that is rugulose or brush-like; the chromatographic surfaces of the monolith are non-porous; the monolith has channels sufficiently large for convective flow of the mobile phase; the monolith is formed from a polymerization mixture including underivatized styrene, a crosslinking agent, and a porogen, wherein the porogen includes tetrahydrofuran. The monolith can be incorporated into a miniaturized chromatography system which can be coupled to a mass spectrometer for on-line separation and mass determination of single- or double-stranded polynucleotides.

摘要翻译： 基于毛细管整体柱的方法和装置，优选由未衍生的聚（苯乙烯 - 二乙烯基苯）整料组成，用于通过离子对 - 反相 - 高性能色谱法（IP-RP-HPLC）分离多核苷酸的混合物。 在该方法和装置的各个方面，整料的特征在于以下一个或多个：整料包含在毛细管内; 通过在管的内壁上共价连接将整料固定化; 管没有保留玻璃料; 该单块的特征在于每米具有超过10,000理论塔板，优选地每平方米超过20万理论塔板; 该方法使用无EDTA的流动相; 整料具有红褐色或刷状的表面形态; 整料的色谱表面是无孔的; 整体结构具有足够大的流动相的对流流动通道; 整料由包含未衍生的苯乙烯，交联剂和致孔剂的聚合混合物形成，其中致孔剂包括四氢呋喃。 整料可以并入小型化的色谱系统中，该系统可与质谱联用，用于单链或双链多核苷酸的在线分离和质量测定。

公开(公告)号：US06372130B1

公开(公告)日：2002-04-16

申请号：US09493734

申请日：2000-01-28

申请人： Douglas T. Gjerde ,  Paul D. Taylor

发明人： Douglas T. Gjerde ,  Paul D. Taylor

IPC分类号： B01D1508

CPC分类号： B01J20/287 ,  B01D15/325 ,  B01D15/366 ,  B01J20/28042 ,  B01J20/3227 ,  B01J20/328 ,  C12N15/1006

摘要： Nonporous beads having an average diameter of about 0.5-100 microns are suitable for chromatographic separation of mixtures of polynucleotides when the beads comprise a nonporous particle which are coated with a polymer or which have substantially all surface substrate groups endcapped with a non-polar hydrocarbon or substituted hydrocarbon group. The beads provide efficient separation of polynucleotides using Matched Ion Polynucleotide Chromatography.

摘要翻译： 平均直径约为0.5-100微米的无孔珠粒适用于多核苷酸混合物的色谱分离，当珠粒包含用聚合物涂覆的无孔颗粒或具有基本上全部被非极性烃封端的表面底物基团时， 取代的烃基。 珠使用匹配离子多核苷酸色谱法提供多核苷酸的有效分离。

公开(公告)号：US06355791B1

公开(公告)日：2002-03-12

申请号：US09562069

申请日：2000-05-01

申请人： Douglas T. Gjerde ,  Paul D. Taylor ,  Robert M. Haefele

发明人： Douglas T. Gjerde ,  Paul D. Taylor ,  Robert M. Haefele

IPC分类号： C07H2104

CPC分类号： B01D15/366 ,  B01D15/20 ,  B01J20/285 ,  C08F8/02 ,  C08F8/30 ,  C08F212/08 ,  C08F230/08 ,  C12N15/101

摘要： Non-polar polymeric separation media, such as beads or monoliths, are suitable for chromatographic separation of mixtures of polynucleotides when the surface of the media are unsubstituted or substituted with a hydrocarbon group having from one to 1,000,000 carbons and when the surfaces are substantially free from multivalent cation contamination. The polymeric media provide efficient separation of polynucleotides using Matched ion Polynucleotide Chromatography. Methods for maintaining and storing the polymeric media include treatment with multivalent cation binding agents.

摘要翻译： 非极性聚合物分离介质，例如珠粒或整料，适用于当介质的表面未被取代或被具有1至1,000,000个碳原子的烃基取代时，多个核苷酸混合物的色谱分离，并且当表面基本上不含 多价阳离子污染 聚合物介质使用匹配离子多核苷酸色谱法提供多核苷酸的有效分离。 维持和储存聚合物介质的方法包括用多价阳离子结合剂进行处理。

公开(公告)号：US20010042714A1

公开(公告)日：2001-11-22

申请号：US09753856

申请日：2001-01-02

发明人： Douglas T. Gjerde ,  David P. Hornby ,  Christopher P. Hanna ,  Alexander I. Kuklin ,  Robert M. Haefele ,  Paul D. Taylor

IPC分类号： B01D015/08

CPC分类号： B01D15/366 ,  C12N15/101

摘要： A Matched Ion Polynucleotide Chromatography method and system for size-based segregation of a mixture of RNA molecules. The method includes applying the mixture to a polymeric separation medium having non-polar surfaces and eluting the RNA molecules with a mobile phase which includes counterion reagent and an organic component. The preferred surfaces are characterized by being substantially free from multivalent cations which are free to interfere with RNA segregation. The elution is preferably performed at a temperature sufficient to denature the RNA. The method can be used in segregating RNA molecules having lengths in the range of about 100 to 20,000 nucleotides. Improved segregation is obtained using a chromatography column having an ID greater than about 5 mm. Examples of separation media include beads and monolithic columns.

摘要翻译： 匹配的离子多核苷酸色谱法和基于尺寸的RNA分子混合物分离的系统。 该方法包括将混合物施加到具有非极性表面的聚合物分离介质中，并用包含抗衡离子试剂和有机组分的流动相洗脱RNA分子。 优选的表面的特征在于基本上不含可以干扰RNA分离的多价阳离子。 洗脱优选在足以使RNA变性的温度下进行。 该方法可用于分离长度在约100至20,000个核苷酸范围内的RNA分子。 使用ID大于约5mm的色谱柱获得改进的分离。 分离介质的实例包括珠粒和整体柱。