公开(公告)号：US07691635B2

公开(公告)日：2010-04-06

申请号：US10590973

申请日：2005-03-24

申请人： Ali Laayoun ,  Eloy Bernal-Mendez

发明人： Ali Laayoun ,  Eloy Bernal-Mendez

IPC分类号： G01N37/00 ,  G01N33/48 ,  C07D239/70 ,  A61K31/4188

CPC分类号： C07D495/04 ,  Y10T436/13 ,  Y10T436/143333

摘要： The present invention relates to a temperature-stable labeling reagent of formula (0): in which: R1 represents H or an alkyl, aryl or substituted aryl group, R2 represents a detectable marker or at least two detectable markers interlinked by at least one multimeric structure, L is a linker arm comprising a linear chain of at least two covalent bonds and n is an integer equal to 0 or 1, R3 and R4 represent, independently of one another: H, NO2, Cl, Br, F, I, R2-(L)n-Y—X—, OR, SR, NR2, R, NHCOR, CONHR, COOR, —CO—NH—(CH2)3—(O—CH2—CH2)3—CH2—NH—R2, —CO—NH—(CH2)3—(O—CH2—CH2)4—CH2—NH—R2 with R=alkyl or aryl, A is a linker arm comprising at least one covalent double bond enabling the conjugation of the diazo function with the aromatic ring and u is an integer between 0 and 2, preferably 0 or 1, —Y—X— represents —CONH—, —NHCO—, —CH2O—, —CH2S—, —Z— represents —NH—, —NHCO—, —CONH— or —O—, m is an integer between 1 and 10, preferably between 1 and 3, and p is an integer between 1 and 10, preferably between 1 and 3. The present invention also describes a method for the synthesis of said labels and also applications for the labeling of biological molecules, in particular of nucleic acids, with a labeling reagent bearing the diazomethyl function.The invention is particularly suitable for use in the field of diagnostics.

摘要翻译： 本发明涉及式（O）的温度稳定的标记试剂：其中：R 1表示H或烷基，芳基或取代的芳基，R 2表示可检测标记或至少两个可检测标记，其与至少一个多聚体 结构，L是包含至少两个共价键的直链的连接臂，n是等于0或1的整数，R 3和R 4彼此独立地表示：H，NO 2，Cl，Br，F，I， R 2 - （L）n Y-X，OR，SR，NR 2，R，NHCOR，CONHR，COOR，-CO-NH-（CH 2）3 - （O-CH 2 -CH 2）3 -CH 2 -NH-R 2， CO-NH-（CH 2）3 - （O-CH 2 -CH 2）4 -CH 2 -NH-R 2，其中R =烷基或芳基，A是连接臂，其包含至少一个共价双键，使得重氮官能团与 芳环和u是0和2之间的整数，优选0或1，-Y-X-表示-CONH - ， - NHCO - ， - CH 2 O - ， - CH 2 S - ， - Z-表示-NH-，-NHCO - ， - CONH-或-O-，m是1 a之间的整数 nd 10，优选在1和3之间，p是1至10之间，优选1至3之间的整数。本发明还描述了合成所述标记的方法以及用于标记生物分子的应用，特别是 的核酸，具有带有重氮甲基功能的标记试剂。 本发明特别适用于诊断领域。

公开(公告)号：US08663923B2

公开(公告)日：2014-03-04

申请号：US12996457

申请日：2009-07-03

申请人： Ali Laayoun ,  Eloy Bernal Mendez

发明人： Ali Laayoun ,  Eloy Bernal Mendez

IPC分类号： C12Q1/68 ,  C07H21/04

CPC分类号： C12Q1/703 ,  C12Q1/6818 ,  C12Q1/701 ,  C12Q2525/307 ,  C12Q2525/301

摘要： A nucleotide probe permitting the detection of nucleic acids and constituted of a labeled nucleotide strand having three fragments: a first fragment having a first closing sequence, a second fragment, all or part of which has a recognition sequence for the molecular recognition of the target nucleic acid and a third fragment having a second closing sequence, and at least two markers, one of the ends of the strand of the detection probe being free of any marker, in which, when the two closing sequences are hybridized together, the detection probe has a completely circular shape, the closing sequences thus maintaining the probe in a conformation that cannot be detected in the absence of the target nucleic acid.

摘要翻译： 允许检测核酸并由具有三个片段的标记的核苷酸链构成的核苷酸探针：具有第一封闭序列的第一片段，其全部或部分具有用于靶核酸的分子识别的识别序列的第二片段 酸和具有第二闭合序列的第三片段和至少两个标记，所述检测探针的链的末端之一没有任何标记，其中当两个封闭序列杂交在一起时，检测探针具有 完全圆形的形状，因此关闭序列因此将探针保持在不存在靶核酸的情况下不能被检测的构象。

公开(公告)号：US20110104762A1

公开(公告)日：2011-05-05

申请号：US12926962

申请日：2010-12-20

申请人： Eloy Bernal-Mendez ,  Ali Laayoun ,  Alain Laurent

发明人： Eloy Bernal-Mendez ,  Ali Laayoun ,  Alain Laurent

IPC分类号： C12P19/34

CPC分类号： C12Q1/6818 ,  C12Q2565/1015 ,  C12Q2525/301 ,  C12Q2525/125 ,  C12Q2525/313

摘要： The present invention relates to a detection probe acting by molecular recognition of a target sequence, comprising successively in the 5′-3′ direction: a first nucleotide segment comprising a sterically hindering structure at its 5′ end, and a second nucleotide segment, complementary to the target sequence.

摘要翻译： 本发明涉及通过靶序列的分子识别作用的检测探针，其连续包含在5'-3'方向：包含在其5'端的空间阻碍结构的第一核苷酸片段和第二核苷酸片段，互补 到目标序列。

公开(公告)号：US20090311688A1

公开(公告)日：2009-12-17

申请号：US12310556

申请日：2007-09-26

申请人： Eloy Bernal-Mendez ,  Ali Laayoun ,  Alain Laurent

发明人： Eloy Bernal-Mendez ,  Ali Laayoun ,  Alain Laurent

IPC分类号： C12Q1/68 ,  C07H21/04 ,  C12N9/99

CPC分类号： C12Q1/6818 ,  C12Q2563/113

摘要： The present invention relates to a labeled oligonucleotide comprising a first nucleotide segment and a second nucleotide segment, complementary to a target sequence, characterized in that it comprises a fluorophore, a quencher and at least one alpha-anomeric nucleoside. The invention also relates to the use of such an oligonucleotide and also to a process using such an oligonucleotide.

摘要翻译： 本发明涉及包含与靶序列互补的第一核苷酸区段和第二核苷酸区段的标记寡核苷酸，其特征在于其包含荧光团，猝灭剂和至少一种α-异头核苷。 本发明还涉及这种寡核苷酸的用途，也涉及使用这种寡核苷酸的方法。

公开(公告)号：US20090215188A1

公开(公告)日：2009-08-27

申请号：US11919668

申请日：2006-05-31

申请人： Eloy Bernal-Mendez ,  Ali Laayoun ,  Lionel Menou

发明人： Eloy Bernal-Mendez ,  Ali Laayoun ,  Lionel Menou

IPC分类号： G01N33/48 ,  C07H1/00

CPC分类号： G01N33/58 ,  G01N33/532 ,  G01N33/54353 ,  Y10T436/11 ,  Y10T436/143333

摘要： The present invention relates to a method for labeling biological molecules of interest contained in a biological sample, consisting in: a) providing a reaction vessel, b) immobilizing capture molecules, which are capable of binding a label of the biological molecules of interest, on a solid support of the vessel, c) introducing the biological sample but also at least one label of the biological molecules of interest into said vessel and optionally any ingredient required for labeling or prelabeling the molecules of interest, d) incubating the vessel content and immobilizing the label which is not reacted with the molecules of interest by binding to the capture molecules, and e) using the labeled molecules of interest for subsequent steps. A method for treating a biological sample is also disclosed.Said invention is preferably used in a manual or automated method for purifying nucleic acids.

摘要翻译： 本发明涉及用于标记生物样品中所含感兴趣生物分子的方法，其包括：a）提供反应容器，b）使能够将感兴趣的生物分子的标记结合的捕获分子固定在 所述容器的固体支持物，c）将所述生物学样品引入所述生物样品，还将所述生物分子的至少一个标记物引入所述容器，以及任选的任何标记或预标记目的分子所需的成分，d）孵育所述容器内容物并固定 通过结合捕获分子不与感兴趣的分子反应的标记，和e）使用目的标记的分子进行后续步骤。 还公开了一种用于处理生物样品的方法。 所述发明优选以用于纯化核酸的手动或自动化方法使用。

公开(公告)号：US08637322B2

公开(公告)日：2014-01-28

申请号：US11919668

申请日：2006-05-31

申请人： Eloy Bernal-Mendez ,  Ali Laayoun ,  Lionel Menou

发明人： Eloy Bernal-Mendez ,  Ali Laayoun ,  Lionel Menou

IPC分类号： G01N33/48 ,  C07H1/00

CPC分类号： G01N33/58 ,  G01N33/532 ,  G01N33/54353 ,  Y10T436/11 ,  Y10T436/143333

摘要： The present invention relates to a method for labeling biological molecules of interest contained in a biological sample, consisting in: a) providing a reaction vessel, b) immobilizing capture molecules, which are capable of binding a label of the biological molecules of interest, on a solid support of the vessel, c) introducing the biological sample but also at least one label of the biological molecules of interest into said vessel and optionally any ingredient required for labeling or prelabeling the molecules of interest, d) incubating the vessel content and immobilizing the label which is not reacted with the molecules of interest by binding to the capture molecules, and e) using the labeled molecules of interest for subsequent steps. A method for treating a biological sample is also disclosed.Said invention is preferably used in a manual or automated method for purifying nucleic acids.

摘要翻译： 本发明涉及用于标记生物样品中所含感兴趣生物分子的方法，其包括：a）提供反应容器，b）使能够将感兴趣的生物分子的标记结合的捕获分子固定在 所述容器的固体支持物，c）将所述生物学样品引入所述生物样品，还将所述生物分子的至少一个标记物引入所述容器，以及任选的任何标记或预标记目的分子所需的成分，d）孵育所述容器内容物并固定 通过结合捕获分子不与感兴趣的分子反应的标记，和e）使用目的标记的分子进行后续步骤。 还公开了一种用于处理生物样品的方法。 所述发明优选以用于纯化核酸的手动或自动化方法使用。

公开(公告)号：US08158345B2

公开(公告)日：2012-04-17

申请号：US12310556

申请日：2007-09-26

申请人： Eloy Bernal-Mendez ,  Ali Laayoun ,  Alain Laurent

发明人： Eloy Bernal-Mendez ,  Ali Laayoun ,  Alain Laurent

IPC分类号： C12Q1/68 ,  C07H21/02 ,  C07H21/04

CPC分类号： C12Q1/6818 ,  C12Q2563/113

摘要： The present invention relates to a labeled oligonucleotide comprising a first nucleotide segment and a second nucleotide segment, complementary to a target sequence, a fluorophore, a quencher and at least one alpha-anomeric nucleoside. The invention also relates to the use of such an oligonucleotide and also to a process using such an oligonucleotide.

摘要翻译： 本发明涉及包含与靶序列，荧光团，猝灭剂和至少一种α-异头核苷酸互补的第一核苷酸区段和第二核苷酸区段的标记寡核苷酸。 本发明还涉及这种寡核苷酸的用途，也涉及使用这种寡核苷酸的方法。

公开(公告)号：US20110091898A1

公开(公告)日：2011-04-21

申请号：US12996457

申请日：2009-07-03

申请人： Ali Laayoun ,  Eloy Bernal Mendez

发明人： Ali Laayoun ,  Eloy Bernal Mendez

IPC分类号： C12Q1/68 ,  C07H21/04

CPC分类号： C12Q1/703 ,  C12Q1/6818 ,  C12Q1/701 ,  C12Q2525/307 ,  C12Q2525/301

摘要： A nucleotide probe permitting the detection of nucleic acids and constituted of a labeled nucleotide strand having three fragments: a first fragment having a first closing sequence, a second fragment, all or part of which has a recognition sequence for the molecular recognition of the target nucleic acid and a third fragment having a second closing sequence, and at least two markers, one of the ends of the strand of the detection probe being free of any marker, in which, when the two closing sequences are hybridized together, the detection probe has a completely circular shape, the closing sequences thus maintaining the probe in a conformation that cannot be detected in the absence of the target nucleic acid.

摘要翻译： 允许检测核酸并由具有三个片段的标记的核苷酸链构成的核苷酸探针：具有第一封闭序列的第一片段，其全部或部分具有用于靶核酸的分子识别的识别序列的第二片段 酸和具有第二闭合序列的第三片段和至少两个标记，所述检测探针的链的末端之一没有任何标记，其中当两个封闭序列杂交在一起时，检测探针具有 完全圆形的形状，因此关闭序列因此将探针保持在不存在靶核酸的情况下不能被检测的构象。

公开(公告)号：US20090117575A1

公开(公告)日：2009-05-07

申请号：US12225693

申请日：2007-04-23

申请人： Eloy Bernal-Mendez ,  Ali Laayoun ,  Alain Laurent

发明人： Eloy Bernal-Mendez ,  Ali Laayoun ,  Alain Laurent

IPC分类号： C12Q1/68 ,  C07H21/04 ,  C12N9/99

CPC分类号： C12Q1/6818 ,  C12Q2565/1015 ,  C12Q2525/301 ,  C12Q2525/125 ,  C12Q2525/313

摘要： The present invention relates to a detection probe acting by molecular recognition of a target sequence, comprising successively in the 5′-3′ direction: a first nucleotide segment comprising a sterically hindering structure at its 5′ end, and a second nucleotide segment, complementary to the target sequence.

摘要翻译： 本发明涉及通过靶序列的分子识别作用的检测探针，其连续包含在5'-3'方向：包含在其5'端的空间阻碍结构的第一核苷酸片段和第二核苷酸片段，互补 到目标序列。

公开(公告)号：US07338805B2

公开(公告)日：2008-03-04

申请号：US10137454

申请日：2002-05-03

申请人： Cecile Bourget ,  Jean Lhomme ,  Ali Laayoun ,  Mitsuharu Kotera ,  Emmanuelle Trevisiol ,  Lionel Menou ,  Eloy Bernal Mendez

发明人： Cecile Bourget ,  Jean Lhomme ,  Ali Laayoun ,  Mitsuharu Kotera ,  Emmanuelle Trevisiol ,  Lionel Menou ,  Eloy Bernal Mendez

IPC分类号： G01N37/00 ,  G01N33/48

CPC分类号： C07D209/08 ,  C07C245/14 ,  C07C245/18 ,  C07D311/90 ,  C07D495/04 ,  Y10T436/13 ,  Y10T436/143333

摘要： The present invention relates to a temperature-stable labeling reagent of formula: in which R1 represents H or an alkyl, aryl or substituted aryl group, R2 represents a detectable marker or at least two detectable markers linked together by at least one multimeric structure, L is a linking arm containing a linear succession of at least two covalent bonds and n an integer equal to 0 or 1, R3 and R4 represent independently of each other: H, NO2, Cl, Br, F, I, R2-(L)n-Y—X—, OR, SR, NR2, R, NHCOR, CONHR, COOR with R=alkyl or aryl, A is a linking arm containing at least one covalent double bond allowing conjugation of the diazo functional group with the aromatic ring and u is an integer between 0 and 2, preferably equal to 0 or 1, and —Y—X— represents —CONH—, —NHCO—, —CH2O—, —CH2S—. The present invention also describes a method for synthesizing said markers as well as applications for labeling biological molecules, in particular nucleic acids, with a labeling reagent carrying the diazomethyl functional group.The invention finds a preferred application in the field of diagnosis.

摘要翻译： 本发明涉及下式的温度稳定的标记试剂：其中R 1表示H或烷基，芳基或取代的芳基，R 2表示可检测标记 或至少两个通过至少一个多聚体结构连接在一起的可检测标记，L是含有至少两个共价键的线性连续并且n为等于0或1的整数的连接臂，R 3和 R 4彼此独立地表示：H，NO 2，Cl，Br，F，I，R 2 - （L） _{O-，-CH 2 S-。 本发明还描述了一种用于合成所述标记的方法以及用携带重氮甲基官能团的标记试剂标记生物分子，特别是核酸的应用。 本发明在诊断领域找到优选的应用。}