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    • 1. 发明授权
    • Nucleic acid isolation
    • 核酸分离
    • US06737235B1
    • 2004-05-18
    • US08945731
    • 1997-11-10
    • Philippe CrosAbdelhamid ElaissariClaude MabilatChristian PichotMarc RodrigueLise Santoro
    • Philippe CrosAbdelhamid ElaissariClaude MabilatChristian PichotMarc RodrigueLise Santoro
    • C07H2104
    • C12N15/101C12N15/1006C12Q1/6806
    • A method for aqueous phase nucleic acid isolation from a sample, comprising a step of nucleic acid adsorption on a particulate substrate, is disclosed. The method comprises an adsorption reagent preparation step (a) that includes a sol consisting of a aqueous continuous phase and a dispersed particulate substrate phase including a functionalized particulate polymer prepared by polymerizing (1) a first water-soluble acrylamide or acrylamide derivative monomer, (2) at least one cross-linking agent and (3) at least one second water-soluble, cationic and functional monomer, said polymer having a predetermined lower critical solubility temperature (LCST) of 25-45° C.; a contact step (b) wherein the adsorption reagent is contacted with the sample containing the nucleic acid; an adsorption step (c) wherein, to carry out the contact step (b), at least one parameter is selected for the reaction medium, said parameters being a pH no higher than 7, an ionic strength no higher than 10−2 M, and a temperature lower than the polymer LCST; a separation step (d) wherein the dispersed phase is separated from the continuous phase, optionally after it has been observed that adsorption has occurred; and a desorption step (e) wherein the nucleic acid is desorbed from the particulate substrate by increasing the ionic strength until an ionic strength higher than 10−2 M is achieved.
    • 公开了一种从样品中分离水相核酸的方法,包括在颗粒基质上的核酸吸附步骤。 该方法包括一种吸附剂制备步骤(a),其包括由水性连续相组成的溶胶和包含通过聚合(1)第一水溶性丙烯酰胺或丙烯酰胺衍生物单体制备的官能化颗粒聚合物的分散颗粒基质相( 2)至少一种交联剂和(3)至少一种第二水溶性阳离子和功能性单体,所述聚合物具有25-45℃的预定的较低临界溶解度温度(LCST)。 接触步骤(b),其中所述吸附剂与含有所述核酸的样品接触; 吸附步骤(c)其中,为了进行接触步骤(b),为反应介质选择至少一个参数,所述参数为不高于7的pH,不高于10 -2的离子强度 M,温度低于聚合物LCST; 分离步骤(d),其中分散相与连续相分离,任选地在已经观察到发生吸附之后; 和解吸步骤(e),其中通过增加离子强度直到达到高于10μM的离子强度,从核粒子脱离。
    • 8. 发明授权
    • Magnetic particles, method for obtaining same and uses for separating molecules
    • 磁性粒子,其获得方法和用于分离分子的方法
    • US06521341B1
    • 2003-02-18
    • US09582352
    • 2000-09-21
    • Abdelhamid ElaissariChristian PichotBernard Mandrand
    • Abdelhamid ElaissariChristian PichotBernard Mandrand
    • G11B516
    • H01F1/111B03C1/01G01N33/5434G01N2446/20G01N2446/86Y10S428/90Y10T428/2991Y10T428/2998
    • The invention concerns magnetic and heat-sensitive particles having a predetermined size between 0.05 and 10 &mgr;m, each particle comprising: an inner composite core consisting of a solid organic or inorganic particle, containing within it a magnetic filler; an outer coati based on a polymer capable of interacting with at least a biological molecule, the outer polymer is heat-sensitive and has a predetermined low critical solubility temperature (LCST) ranging between 10 and 100° C. and preferably between 20 and 60° C. The invention also concerns methods for obtaining said particles and their uses. The particles are characterized in that there is an intermediate layer between the inner core and the outer layer isolating said inner core magnetic filler with respect to said functionalized outer layer. Said invention is particularly useful for separating proteins and/or nucleic acids.
    • 本发明涉及具有0.05-10μm之间的预定尺寸的磁性和热敏颗粒,每个颗粒包括:由固体有机或无机颗粒组成的内部复合芯,其中含有磁性填料; 基于能够与至少一种生物分子相互作用的聚合物的外涂层,外部聚合物是热敏感的,并且具有在10和100℃之间,优选在20和60℃之间的预定的低临界溶解温度(LCST) 本发明还涉及获得所述颗粒及其用途的方法。 颗粒的特征在于,在内芯和外层之间存在相对于所述官能化外层隔离所述内芯磁性填料的中间层。 所述发明对于分离蛋白质和/或核酸特别有用。
    • 9. 发明授权
    • Reagent and method for the detection of a nucleotide sequence with
signal amplification
    • 用于检测具有信号放大的核苷酸序列的试剂和方法
    • US6017707A
    • 2000-01-25
    • US870730
    • 1997-06-06
    • Bernard MandrandPhilippe CrosThierry DelairMarie-Helene CharlesMarie-Noelle EroutChristian Pichot
    • Bernard MandrandPhilippe CrosThierry DelairMarie-Helene CharlesMarie-Noelle EroutChristian Pichot
    • C08G81/02C12Q1/68C07H21/02C07H21/04C12N15/00
    • C12Q1/6813C08G81/02C12Q1/682
    • Methods for the detection of a nucleotide sequence of interest comprising at least one nucleotide probe marked with a tracer. The methods include a reagent essentially comprising a linear backbone copolymer having lateral substituents, whose chain consists of a first type of repetitive unit and at least one other type of repetitive unit, in which at least one part of the units of the first type have a lateral substituent comprising a nucleotide unit, such a lateral substituent not being present on the other types of units. Each of said nucleotide units, all of which are identical, comprise at least one nucleotide sequence capable of hybridizing with said sequence of interest and nucleotide sequence capable of hybridizing with a probe, the reagent containing on average more than two of said nucleotide units, in molar equivalents, per mole of polymer. Such a reagent enables signal amplification to be obtained, and therefor lowers the sensitivity threshold. Application, in particular, in the production of tests for the detection of pathogenic organisms, or in the diagnosis of genetic diseases.
    • 检测目的核苷酸序列的方法包括用示踪剂标记的至少一个核苷酸探针。 所述方法包括基本上包含具有侧向取代基的线性主链共聚物的试剂,其链由第一类型的重复单元和至少一种其它类型的重复单元组成,其中第一类型的单元的至少一部分具有 包含核苷酸单元的侧向取代基,这样的侧向取代基不存在于其它类型的单元上。 所述全部相同的所述核苷酸单元包含至少一个能够与所述感兴趣序列杂交的核苷酸序列和能够与探针杂交的核苷酸序列,所述试剂平均包含两个以上的所述核苷酸单元, 摩尔当量,每摩尔聚合物。 这样的试剂能够获得信号放大,从而降低灵敏度阈值。 特别是应用于生产用于检测病原体的检测或遗传疾病的诊断。
    • 10. 发明授权
    • Reagent and method for the detection of a nucleotide sequence with
signal amplification
    • 用于检测具有信号放大的核苷酸序列的试剂和方法
    • US5695936A
    • 1997-12-09
    • US433505
    • 1995-06-14
    • Bernard MandrandPhilippe CrosThierry DelairMarie-Helene CharlesMarie-Noelle EroutChristian Pichot
    • Bernard MandrandPhilippe CrosThierry DelairMarie-Helene CharlesMarie-Noelle EroutChristian Pichot
    • C08G81/02C12Q1/68
    • C12Q1/6813C08G81/02C12Q1/682
    • Methods for the detection of a nucleotide sequence of interest comprising at least one nucleotide probe marked with a tracer. The methods comprise use of a reagent essentially comprising a linear backbone copolymer having lateral substituents, whose chain consists of a first type of repetitive unit and at least one other type of repetitive unit, in which at least one part of the units of the first type have a lateral substituent comprising a nucleotide unit, such a lateral substituent not being present on the other types of units. Each of said nucleotide units, all of which are identical, comprise at least one nucleotide sequence capable of hybridizing with said sequence of interest and nucleotide sequence capable of hybridizing with a probe, the reagent containing on average more than two of said nucleotide units, in molar equivalents, per mole of polymer. Such a reagent enables signal amplification to be obtained, and therefor lowers the sensitivity threshold. Application, in particular, in the production of tests for the detection of pathogenic organisms, or in the diagnosis of genetic diseases.
    • PCT No.PCT / FR94 / 01084第 371 1995年6月14日第 102(e)1995年6月14日PCT PCT 1994年9月15日PCT公布。 WO95 / 08000 PCT出版物 1995年3月23日的方法检测目标核苷酸序列的方法包括用示踪剂标记的至少一个核苷酸探针。 所述方法包括使用基本上包含具有侧向取代基的线性主链共聚物的试剂,其链由第一类型的重复单元和至少一种其它类型的重复单元组成,其中第一类型的单元的至少一部分 具有包含核苷酸单元的侧向取代基,这样的侧向取代基不存在于其它类型的单元上。 所述全部相同的所述核苷酸单元包含至少一个能够与所述感兴趣序列杂交的核苷酸序列和能够与探针杂交的核苷酸序列,所述试剂平均包含两个以上的所述核苷酸单元, 摩尔当量,每摩尔聚合物。 这样的试剂能够获得信号放大,从而降低灵敏度阈值。 特别是应用于生产用于检测病原体的检测或遗传疾病的诊断。