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    Solid phases optimized for chemiluminescent detection
    2.
    发明授权
    Solid phases optimized for chemiluminescent detection 有权
    为化学发光检测优化的固相

    公开(公告)号：US07368296B2

    公开(公告)日：2008-05-06

    申请号：US10046730

    申请日：2002-01-17

    Applicant: Brooks Edwards ,  Timothy G. Geiser ,  Steven M. Menchen ,  Alison L. Sparks ,  John C. Voyta

    Inventor： Brooks Edwards ,  Timothy G. Geiser ,  Steven M. Menchen ,  Alison L. Sparks ,  John C. Voyta

    IPC: G01N33/545 ,  G01N33/533 ,  C07C211/62

    CPC classification number: C12Q1/6823 ,  B01J2219/00659 ,  B01J2219/00702 ,  B01J2219/00722 ,  B01J2219/00725 ,  C40B40/06 ,  C40B40/10 ,  C40B50/14 ,  G01N33/54393 ,  Y10S436/805 ,  Y10S436/823

    Abstract: Solid supports for chemiluminescent assays are provided. The solid support includes a plurality of probes covalently or physically attached to the support surface and a chemiluminescent enhancing moiety incorporated onto the surface or into the bulk of the support. The solid support can be a multi-layered support including an upper probe binding layer (e.g., an azlactone polymer layer or porous functional polyamide layer) adjacent to a cationic microgel layer. The azlactone-functional polymer can be a copolymer of dimethylacrylamide and vinylazlactone crosslinked with ethylenediamine. The cationic microgel layer can be a cross-linked quaternary onium salt containing polymer. A method and a kit for conducting chemiluminescent assays using the solid supports is also provided. The kit comprises a dioxetane substrate, a biopolymer probe-enzyme complex, and a solid support. The solid support can be an azlactone functional polymer layer adjacent to a cationic microgel layer; a porous polyamide functional layer adjacent to a cationic microgel layer; or a quaternized azlactone functional polymer layer.

    Abstract translation: 提供了化学发光测定的固体支持物。 固体支持物包括共价或物理连接到支持物表面上的多个探针和结合到表面或载体的大部分中的化学发光增强部分。 固体支持物可以是包含邻近阳离子微凝胶层的上探针结合层（例如，吖内酯聚合物层或多孔功能性聚酰胺层）的多层载体。 吖内酯官能聚合物可以是与乙二胺交联的二甲基丙烯酰胺和乙烯基内酯的共聚物。 阳离子微凝胶层可以是交联的含季铵盐的聚合物。 还提供了使用固体支持物进行化学发光测定的方法和试剂盒。 试剂盒包括二氧杂环丁烷底物，生物聚合物探针 - 酶复合物和固体支持物。 固体支持物可以是与阳离子微凝胶层相邻的吖内酯官能聚合物层; 与阳离子微凝胶层相邻的多孔聚酰胺官能层; 或季铵化的吖内酯官能聚合物层。

    Simultaneous generation of multiple chemiluminescent signals on solid supports
    3.
    发明申请
    Simultaneous generation of multiple chemiluminescent signals on solid supports 审中-公开
    在固体支持物上同时产生多种化学发光信号

    公开(公告)号：US20050026151A1

    公开(公告)日：2005-02-03

    申请号：US10620333

    申请日：2003-07-17

    Applicant: John Voyta ,  Robert Smith ,  Gary Schroth ,  Alison Sparks ,  Brooks Edwards

    Inventor： John Voyta ,  Robert Smith ,  Gary Schroth ,  Alison Sparks ,  Brooks Edwards

    IPC: G01N33/58 ,  C12Q1/68

    CPC classification number: G01N33/581

    Abstract: A chemiluminescent assay to determine the presence and/or amount of one or more labeled target molecules in a sample is described in which the surface layer of a solid support is contacted with a composition comprising first and second chemiluminescent substrates capable of being activated by first and second enzymes, respectively. A plurality of probes are disposed on the surface layer in discrete areas. At least some of the probes are bound to a first enzyme conjugate comprising the first enzyme and at least some of the probes are bound to a second enzyme conjugate comprising the second enzyme. The resulting chemiluminescent signals are then detected. The method can be used to compare two biological samples (e.g., mRNA populations from different cells) on the same support surface or to provide a chemiluminescent control signal for normalizing chemiluminescent assay data from a biological sample.

    Abstract translation: 描述了用于确定样品中一种或多种标记的靶分子的存在和/或量的化学发光测定法，其中固体支持物的表面层与包含能够被第一和第二化学发光底物活化的第一和第二化学发光底物的组合物接触， 第二种酶。 多个探针设置在离散区域的表面层上。 至少一些探针与包含第一酶的第一酶结合物结合，并且至少一些探针与包含第二酶的第二酶缀合物结合。 然后检测所得到的化学发光信号。 该方法可用于比较两个生物样品（例如，来自不同细胞的mRNA群体）在相同的支持物表面上，或提供化学发光控制信号，用于从生物样品中归一化化学发光测定数据。

    Dioxetane labeled probes and detection assays employing the same
    4.
    发明授权
    Dioxetane labeled probes and detection assays employing the same 有权
    二氧杂环丁烷标记的探针和使用其的检测测定

    公开(公告)号：US06451531B1

    公开(公告)日：2002-09-17

    申请号：US09340726

    申请日：1999-06-29

    Applicant: Irena Bronstein ,  Brooks Edwards ,  Christopher Martin ,  John Voyta

    Inventor： Irena Bronstein ,  Brooks Edwards ,  Christopher Martin ,  John Voyta

    IPC: C12Q168

    CPC classification number: C07H21/00 ,  C07D321/00 ,  C12Q1/6816 ,  C12Q1/686 ,  Y02P20/55 ,  C12Q2561/101 ,  C12Q2565/113

    Abstract: Probes labeled with 1,2-dioxetane precursors can be employed in a variety of assays. The probes may be nucleic acid, peptide nucleic acid, proteins including enzyme, antibody or antigen, steroid, carbohydrate, drug or non-drug hapten. The probe is provided with a 1,2-dioxetane precursor bound thereto, generally either covalently, or a strong ligand bond. The dioxetane precursor moiety is converted to a bound 1,2-dioxetane by exposure to singlet oxygen. These dioxetane (labels) either spontaneously decompose, or are induced to decompose by an appropriate trigger to release light. The trigger may be a change in pH temperature, or an agent which removes a protective group. Assay formats in which these 1,2-dioxetane labeled probes and referents may be used to include hybridization assays, immuno assays, gel-based assays and Capillary Zone Electrophoresis.

    Abstract translation: 用1,2-二氧杂环丁烷前体标记的探针可用于多种测定。 探针可以是核酸，肽核酸，蛋白质，包括酶，抗体或抗原，类固醇，碳水化合物，药物或非药物半抗原。 探针具有与其结合的1,2-二氧杂环丁烷前体，通常共价或强配体键。 通过暴露于单线态氧将二氧杂环丁烷前体部分转化为结合的1,2-二氧环乙烷。 这些二氧杂环丁烷（标记）会自发分解，或被适当的触发剂诱导分解以释放光。 触发可能是pH温度的变化，或者是除去保护基团的试剂。 这些1,2-二氧杂环丁烷标记的探针和指示物可用于包括杂交测定，免疫测定，基于凝胶的测定和毛细管区域电泳的测定形式。

    Synthesis of 1, 2-dioxetanes and intermediates therefor
    8.
    发明授权
    Synthesis of 1, 2-dioxetanes and intermediates therefor 失效
    1,2-二氧杂环丁烷及其中间体的合成

    公开(公告)号：US5777133A

    公开(公告)日：1998-07-07

    申请号：US791050

    申请日：1997-01-28

    Applicant: Irena Y. Bronstein ,  Brooks Edwards

    Inventor： Irena Y. Bronstein ,  Brooks Edwards

    IPC: C07C43/178 ,  C07C43/196 ,  C07C43/215 ,  C07C69/00 ,  C07D321/00 ,  C07F9/12 ,  C07F9/655 ,  C07F9/6574 ,  C07H15/203 ,  C07H15/26 ,  C12Q1/34 ,  G01N21/76 ,  G01N33/58 ,  C07F9/28

    CPC classification number: C07H15/203 ,  C07C43/1788 ,  C07C43/196 ,  C07C43/215 ,  C07C69/00 ,  C07D321/00 ,  C07F9/12 ,  C07F9/65512 ,  C07F9/65742 ,  C07H15/26 ,  C12Q1/34 ,  G01N21/76 ,  G01N33/581 ,  G01N33/582 ,  C07C2103/00 ,  C07C2103/74

    Abstract: Compounds having the formula: ##STR1## wherein T is a polycycloalkylidene group (e.g., adamant-2-ylidene); R is a C.sub.1-20 alkyl, aralkyl or cycloalkyl group; and Y is a fluorescent chromophore (eg., m-phenylene), produced by reacting a compound having the formula: ##STR2## with an R-ylating agent (e.g., R.sub.2 SO.sub.4) in the presence of an alkali metal alkoxide in a polar aprotic solvent. Also, compounds having the formula: ##STR3## are produced by reacting a compound having the formula: ##STR4## wherein X is an electronegative leaving group (e.g., a halogen anion such as chloride ion) in the presence of a Lewis base (e.g., a trialkyl-amine) dissolved in an aprotic organic solvent (e.g., benzene or toluene). Also, compounds having the formula ##STR5## are produced by reacting a compound of the formula ##STR6## with a tetra-O-acylated-O-hexopyranoside halide, then hydrolyzing off the protective acyl groups. The aforementioned compounds and procedures are useful in the synthesis of enzyme-cleavable 1,2-dioxetane ring systems that can serve as members of a binding pair employed, for example, in chemiluminescent immunoassays, DNA probe assays, and direct assays for an enzyme.

    Abstract translation: 具有下式的化合物：其中T是多环亚烷基（例如，金刚烷-2-亚基）; R是C 1-20烷基，芳烷基或环烷基; Y是荧光发色团（例如，间亚苯基），其通过在极性的碱金属醇盐存在下，使具有下式的化合物与<！ - SIPO 是通过使下式化合物与其中X是电负离子基团（例如卤素阴离子如氯离子）反应的化合物与 溶解在非质子有机溶剂（例如苯或甲苯）中的路易斯碱（例如三烷基胺）。 此外，具有式“IMAGE”的化合物通过使式“IMAGE”的化合物与四-O-酰化的O-六吡喃苷卤化物反应，然后水解掉保护性酰基而产生。 上述化合物和方法可用于合成可切割的1,2-二氧杂环丁烷环系统，其可以用作结合对的成员，例如在化学发光免疫测定，DNA探针测定和酶的直接测定中使用。

    1,2 chemiluminescent dioxetanes of improved performance
    9.
    发明授权
    1,2 chemiluminescent dioxetanes of improved performance 失效
    1,2化学发光二恶烷具有改进的性能

    公开(公告)号：US5679803A

    公开(公告)日：1997-10-21

    申请号：US547372

    申请日：1995-10-25

    Applicant: Irena Bronstein ,  Brooks Edwards

    Inventor： Irena Bronstein ,  Brooks Edwards

    IPC: C07D305/14 ,  C07D321/00 ,  C07F9/06 ,  C07F9/655 ,  C07G3/00 ,  C12Q1/37 ,  G01N33/58 ,  C07F7/02

    CPC classification number: C07D321/00 ,  C07F9/65512 ,  C07G3/00 ,  C12Q1/37 ,  G01N33/582 ,  Y02P20/55 ,  Y10S435/81

    Abstract: A new class of stable dioxetanes bears a polycyclic stabilizing group and aryloxy moiety, the oxygen atom of which is provided with a protective group which can be removed by an enzymatic or chemical trigger admixed with the dioxetane. The polycyclic stabilizing group is preferably spiroadamantane. The group further bears an alkoxy, aryloxy, aralkyloxy or cycloalkyloxymoiety which is partially or completely substituted with halogens, particularly fluorine and chlorine. Proper selection of electron active groups on the stabilizing moiety, the aryl group and the OR group yields enhanced enzyme kinetics, superior light intensity and excellent detection sensitivity in various assays.

    Abstract translation: 一类稳定的二氧杂环丁烷具有多环稳定基团和芳氧基部分，其氧原子具有可以通过与二氧杂环丁烷混合的酶或化学触发剂除去的保护基团。 多环稳定基团优选为四金刚烷。 该基团还具有部分或完全被卤素，特别是氟和氯取代的烷氧基，芳氧基，芳烷氧基或环烷氧基。 稳定部分，芳基和OR基上的电子活性基团的适当选择在各种测定中产生增强的酶动力学，优异的光强度和优异的检测灵敏度。

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