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公开(公告)号：US5149630A
公开(公告)日：1992-09-22
申请号：US157100
申请日：1988-02-09
IPC分类号： C12M1/40 , C12Q1/00 , G01N27/27 , G01N27/30 , G01N27/327 , G01N27/416 , G01N27/48 , G01N33/53 , G01N33/542 , G01N33/543
CPC分类号： G01N33/5438 , Y10S435/81 , Y10S435/817 , Y10S436/806 , Y10S436/808
摘要： An electrochemical specific binding assay of a ligand (e.g., antigen, hapten or antibody) wherein at least one of the components is enzyme-labelled, and which includes the step of determining the extent to which the transfer of electrons between the enzyme substrate and an electrode, associated with the substrate reaction, is perturbed by complex formation or by displacement of any ligand complex relative to unbound enzyme-labelled component.The electron transfer is aided by electron-transfer mediators which can accept electrons from the enzyme and donate them to the electrode or vice versa (e.g. ferrocene) or by electron-transfer promoters which retain the enzyme in close proximity with the electrode without themselves taking up a formal charge.The electrochemical apparatus will typically comprise two or three electrodes, including one working electrode onto which components may advantageously be immobilized.The use of direct electrochemical measurement of the enzyme label avoids the errors and inconvenience of the known indirect measurement techniques.
摘要翻译： 配体（例如抗原，半抗原或抗体）的电化学特异性结合测定，其中至少一种组分是酶标记的，其包括确定酶底物和 与底物反应相关的电极通过复合物形成或通过相对于未结合的酶标记的组分置换任何配位体复合物来扰动。 电子转移辅助于电子转移介质，其可以接受来自酶的电子并将其提供给电极或反之亦然（例如二茂铁）或通过电子转移启动子保留酶极靠近电极而不自身吸收 正式收费。 电化学装置通常包括两个或三个电极，包括一个工作电极，组分可以有利地固定在该工作电极上。 使用酶标记的直接电化学测量避免了已知间接测量技术的错误和不便。
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公开(公告)号：US4945045A
公开(公告)日：1990-07-31
申请号：US212043
申请日：1988-06-23
IPC分类号： G01N27/48 , G01N33/542 , G01N33/543
CPC分类号： G01N33/5438 , G01N27/3271 , Y10S435/817 , Y10S435/975 , Y10S436/806
摘要： A ligand in a sample can be assayed using electrochemical apparatus containing a working electrode and components comprising:(a) the sample,(b) a specific binding partner to the ligand, or a specific binding partner to the ligand and selected from ligand analogues and specific binding partners, and(c) an electron-source or electron-acceptor; at least one of components (b) being labelled with an electron-transfer mediator capable of aiding the transfer of electrons between the electron-source or electron-acceptor and the working electrode by accepting electrons from the electron-source and donating them to the working electrode, or accepting the electrons from the electrode and donating them to the electron-acceptor.The method includes the step of determining whether, and, if desired, the extent to which, the said transfer of electrons is perturbed by at least one of (i) formation of a complex of said ligand with a specific binding partner and (ii) controlled external influence which produces a perturbation of said transfer of electrons as a function of said complex formation.
摘要翻译： 可以使用含有工作电极和组分的电化学装置来测定样品中的配体，所述电化学装置包含：（a）样品，（b）与配体的特异性结合配偶体或配体的特异性结合配偶体，并且选自配体类似物和 特异性结合配偶体，和（c）电子源或电子受体; 组分（b）中的至少一种被电子转移介质标记，该电子传递介质能够通过接受来自电子源的电子并将它们供给到工作中来辅助电子源或电子受体与工作电极之间的电子传递 或者从电极接收电子并将它们提供给电子受体。 该方法包括以下步骤：确定是否和如果需要，所述电子转移的程度被（i）形成所述配体与特异性结合配偶体的复合物和（ii）至少一种所扰动， 控制的外部影响，其产生作为所述络合物形成的函数的所述电子传递的扰动。
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公开(公告)号：US4711245A
公开(公告)日：1987-12-08
申请号：US607699
申请日：1984-05-07
CPC分类号： A61B5/14865 , A61B5/6848 , C12Q1/001 , C12Q1/004 , Y10S435/817
摘要： This invention relates to equipment and methods for detecting the presence of, measuring the amount of, and/or monitoring the level of, one or more selected components in a liquid mixture, employing an electrode sensing system.We have discovered that a class of mediating compounds has extremely useful properties for mediating the transfer of charge between enzyme-catalyzed reactions and electrode surfaces (15) in electrode sensing systems.Specifically, the specification discloses as electrode sensor mediators, organometallic compounds composed of at least two organic rings, each of which has at least two double bonds in a conjugated relationship; a metal atom is in electron-sharing contact with those rings. An enzyme capable of catalyzing a reaction at a rate representative of the selected compound concentration is in contact with an assay mixture, and the mediator compound transfers charge between the enzyme and the conductive surface of the electrode at a rate representative of the enzyme catalyzed reaction rate.
摘要翻译： 本发明涉及使用电极感测系统来检测液体混合物中一种或多种所选组分的存在，测量量和/或监测液位的设备和方法。 我们已经发现，一类介导化合物具有非常有用的性质，用于介导电极感应系统中酶催化反应和电极表面（15）之间的电荷转移。 具体地说，本说明书公开了电极传感器介质，由至少两个有机环组成的有机金属化合物，每个有机环具有至少两个共轭关系的双键; 金属原子与这些环电子共享接触。 能够以代表所选化合物浓度的速率催化反应的酶与测定混合物接触，并且介体化合物以代表酶催化反应速率的速率在酶和电极的导电表面之间转移电荷 。
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公开(公告)号：US4978610A
公开(公告)日：1990-12-18
申请号：US76265
申请日：1987-07-23
IPC分类号： G01N33/532 , G01N33/543
CPC分类号： G01N33/532 , G01N33/54326 , G01N33/5438 , Y10S204/05 , Y10S435/817 , Y10S436/806
摘要： In its broadest aspect, the present invention provides a method of effecting an electrochemical specific-binding assay of a ligand, either qualitatively or quantitatively, in an apparatus comprising at leasts one electrode, in which method a labelled component of the assay medium is, at least in part, magnetically held in the vicinity of the electrode. The electrochemical assay method may include a separation step, whereby bound label may be separated from free label in the assay medium. Preferably the electrochemical assay method will include the step of determining a perturbation in an electrochemical characteristic of components of the assay medium associated with a ligand complexing reaction.Such an assay method is applicable to, for example, antibodies and antigens and suitable labels include redox centers, enzyme labels in the presence of an electron-transfer mediator and electron-transfer mediator labels in the presence of an electron donor or acceptor.
摘要翻译： 在其最广泛的方面，本发明提供了在包括至少一个电极的装置中定性或定量地进行配体的电化学特异性结合测定的方法，其中测定介质的标记组分在 至少部分地磁性地保持在电极附近。 电化学测定方法可以包括分离步骤，其中结合的标记物可以在测定培养基中与游离标记物分离。 优选地，电化学测定方法将包括确定与配体络合反应相关的测定介质的组分的电化学特性的扰动的步骤。 这种测定方法适用于例如抗体和抗原，合适的标记包括氧化还原中心，电子转移介质存在下的酶标记和电子给体或受体存在下的电子转移介质标记。
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公开(公告)号：US4836904A
公开(公告)日：1989-06-06
申请号：US839699
申请日：1986-03-13
IPC分类号： A61B5/145 , A61B5/1473 , A61B5/1486 , G01N27/30 , G01N27/327
CPC分类号： G01N27/308 , G01N27/3277
摘要： An electrode has a graphite surface at least part of which is an edge-plane graphite surface to which Cr(III) complexes are attached by surface C--O groups. The graphite can be polished pyrolytic graphite. The complex coverage can be 5-20%. The complex may comprise proteins, enzyme substrates, specific binding agents, intermediate metabolites or the like, e.g. the protein plastocyanin, so that direct unmediated electrochemistry is possible at the electrode.
摘要翻译： 电极具有石墨表面，其至少一部分是边缘平面石墨表面，Cr（III）配合物通过表面C-O基团连接到该石墨表面。 石墨可以是抛光的热解石墨。 复合覆盖率可以是5-20％。 复合物可以包括蛋白质，酶底物，特异性结合剂，中间体代谢物等。 蛋白质质体蛋白质，使得在电极处可以直接不介导的电化学。
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公开(公告)号：US4820636A
公开(公告)日：1989-04-11
申请号：US832452
申请日：1986-02-21
申请人： Hugh A. O. Hill , Monika J. Green
发明人： Hugh A. O. Hill , Monika J. Green
CPC分类号： G01N33/723 , C12Q1/004 , G01N33/66 , Y10S435/817
摘要： The invention disclosed relates to an electrochemical assay which is particularly concerned with an assay for the presence of, or amount of, glycosylated haemoglobin in a blood sample, but which extends to a general assay for detecting the presence of, monitoring the level of or determining the concentration of compounds containing cis-diol groups, such as glycols, nucleic acid components, sugars, polyols, catechols and glycosylated proteins.When ferrocene boronic acid or a derivative thereof is present free in solution, it can act as a mediator for a wide range of oxido-reductases. However in the presence of, for example, sugars containing the cis-diol structure, the metallocene (such as ferrocene boronic acid) covalently binds to the sugar. When the ferrocene boronic acid is thus bound, it is essentially immobilized and its properties as a mediator and ability to diffuse will be markedly affected.
摘要翻译： 所公开的本发明涉及一种电化学测定法，其特别涉及血液样品中糖基化血红蛋白的存在或量的测定，其扩展到用于检测存在或监测血液样品的水平或确定 含有顺式二醇基团的化合物如二醇，核酸组分，糖，多元醇，邻苯二酚和糖基化蛋白质的浓度。 当二茂铁硼酸或其衍生物在溶液中游离时，可以作为广泛范围氧化还原酶的介质。 然而，在例如含有顺式二醇结构的糖的存在下，金属茂（如二茂铁硼酸）与糖共价结合。 当二茂铁硼酸结合时，其基本上是固定的，其作为介体的性质和扩散能力将受到显着影响。
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7.发明授权
公开(公告)号：US4655885A
公开(公告)日：1987-04-07
申请号：US817570
申请日：1986-01-10
CPC分类号： C12Q1/003
摘要： In a bioelectrochemical process in which the electrons are transferred directly, without use of a mediator, such as a redox dye or cofactor, between an electrode and an electroactive biological material, such as an enzyme or a protein, in either direction, rapid electron transfer has previously been achieved between an electrode and the positively charged protein horse-heart cytochrome c by adding a surface-modifier such as 4,4'-bipyridyl or a derivative thereof.It has now been found possible to promote electron transfer to either a positively or a negatively charged protein using the same surface-modifier for either job, namely a compound of formula ##STR1## wherein: the pyridine ring shown is substituted in the 2-, 3- or 4-position by the (methylene)hydrazinecarbothioamide group shown;R.sup.1 represents hydrogen atom(s) or one or two methyl or ethyl groups in the 2-,3- or 4-position (when the said position is not substituted by the (methylene)hydrazinecarbothioamide group shown);R.sup.2 represents a hydrogen atom or a methyl group;R.sup.3 represents a hydrogen atom or a methyl group; andR.sup.4 represents a hydrogen atom or a methyl group.
摘要翻译： 在生物电化学工艺中，电子直接转移，而不使用电极和电活性生物材料（如酶或蛋白质）之间的任何一个方向，不使用介质（如氧化还原染料或辅因子），快速电子转移 之前已经通过添加表面改性剂如4,4'-联吡啶或其衍生物在电极和带正电荷的蛋白质马 - 心细胞色素c之间实现。 现在已经发现可以使用相同的表面改性剂促进电子转移为正电荷或带负电荷的蛋白质，即任一工作的化合物，即式（1）的化合物，其中：所示的吡啶环在 （亚甲基）肼基硫代酰胺基团的2-，3-或4-位; R1表示2-，3-或4-位的氢原子或一个或两个甲基或乙基（当所述位置未被所示的（亚甲基）肼甲硫基酰胺基团取代时）; R2表示氢原子或甲基; R3表示氢原子或甲基; 并且R 4表示氢原子或甲基。
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公开(公告)号：US4840893A
公开(公告)日：1989-06-20
申请号：US769629
申请日：1985-10-15
申请人： Hugh A. O. Hill , Susan I. Libor
发明人： Hugh A. O. Hill , Susan I. Libor
IPC分类号： C12Q1/00 , C12Q1/54 , C12Q1/68 , G01N33/543 , G01N33/58
CPC分类号： G01N33/581 , C12Q1/00 , C12Q1/004 , C12Q1/54 , C12Q1/6825 , G01N33/5438 , Y10S435/81 , Y10S435/817 , Y10S436/806 , Y10S436/904
摘要： The present invention discloses an assay for nucleic acid which comprises the steps of;(a) providing a probe material comprising;(i) a sequence of nucleic acids complementary to a given target sequence and,(ii) a first ligand chemically linked thereto and capable of a specific binding reaction with an antiligand;(b) contacting the said probe material with an assay system comprising:(i) a suitable mediator, enzyme, substrate system capable of transferring charge to an electrode surface when the enzyme is catalytically active, and;(ii) a second ligand chemically linked to one of said mediator, enzyme or substrate, wherein the second ligand is capable of a competitive binding reaction with the antiligand, and;(iii) the said antiligand, Whereby the said first ligand competes with the said second ligand in a specific binding reaction with the antiligand, and;(c) contacting the above system with a solution suspected of containing the said target sequence whereby the binding of any of the said target sequence present to the probe affects the availability of the first ligand and therefore alters the rate of charge transfer to the electrode.
摘要翻译： PCT No.PCT / GB84 / 00432 Sec。 371日期1985年10月15日第 102（e）日期1985年10月15日PCT提交1984年12月14日PCT公布。 出版物WO85 / 02627 1985年6月20日，本发明公开了一种核酸检测方法，包括以下步骤： （a）提供探针材料，包括： （i）与给定靶序列互补的核酸序列，和（ii）与其结合化学连接且能够与抗配体结合的特异性结合反应的第一配体; （b）使所述探针材料与测定系统接触，所述测定系统包括：（i）当酶是催化活性时能够将电荷转移到电极表面的合适的介体，酶，底物系统; （ii）与所述介体，酶或底物之一化学连接的第二配体，其中所述第二配体能够与所述反配体发生竞争性结合反应;和 （iii）所述抗配糖剂，其中所述第一配体与所述第二配体竞争与所述反配体的特异性结合反应; （c）将上述系统与疑似含有所述靶序列的溶液接触，由此存在的任何所述靶序列与探针的结合影响第一配体的可用性，并因此改变电荷向电极转移的速率。
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公开(公告)号：US4318784A
公开(公告)日：1982-03-09
申请号：US192504
申请日：1980-04-15
申请人： Irving J. Higgins , Hugh A. O. Hill
发明人： Irving J. Higgins , Hugh A. O. Hill
IPC分类号： A61K38/00 , C12M1/40 , C12N9/02 , C12P1/00 , C12P7/02 , C12P7/04 , C12P7/24 , C12R1/01 , C12R1/40 , C25B3/04
摘要： There are provided processes for carrying out enzymatic reactions, especially processes for the oxidative or reductive transformation of organic compounds catalyzed by enzymes, in which the enzymes require a continuing supply of reducing equivalents to regenerate reduced enzyme species in enzymatically active form, and in which the reducing equivalents are derived electrochemically, including electrochemically from molecular hydrogen, the reducing equivalents preferably being passed directly to the enzyme from the cathode of an electrochemical system.
摘要翻译： 提供了用于进行酶促反应的方法，特别是用于由酶催化的有机化合物的氧化还原转化的方法，其中酶需要连续供应还原当量以再生酶活性形式的还原酶物质，并且其中 还原当量是电化学衍生的，包括从分子氢中电化学，还原当量优选从电化学系统的阴极直接传递到酶。
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公开(公告)号：US5509410A
公开(公告)日：1996-04-23
申请号：US281237
申请日：1994-07-27
IPC分类号： A61B5/00 , C12Q1/00 , C12Q1/68 , G01N27/403 , G01N33/487 , G01N33/543
CPC分类号： G01N33/5438 , A61B5/1486 , A61B5/6848 , C12Q1/001 , C12Q1/004 , C12Q1/6825 , G01N27/4035 , A61B2560/0418 , Y10T29/4914 , Y10T29/49153 , Y10T29/49169 , Y10T29/49224
摘要： A sensor system that detects a current representative of a compound in a liquid mixture features a two-electrode strip adapted for releasable attachment to signal readout circuitry. The strip comprises an elongated support (preferably flat) adapted for releasable attachment to the readout circuitry; a first conductor and a second conductor each extend along the support and comprise means for connection to the circuitry. An active electrode, positioned to contact the liquid mixture and the first conductor, comprises a deposit of an enzyme capable of catalyzing a reaction involving the compound and (preferably) an electron mediator, capable of transferring electrons between the enzyme-catalyzed reaction and the first conductor. A reference electrode is positioned to contact the mixture and the second conductor. The system includes circuitry adapted to provide an electrical signal representative of the current. The two-electrode strip is manufactured, e.g., by screen printing an admixture of enzyme, a conductive material, and a mediator as a single layer onto the substrate.
摘要翻译： 检测液体混合物中的化合物的当前代表的传感器系统具有适于可释放地附接到信号读出电路的双电极条。 条带包括适于可释放地附接到读出电路的细长支撑件（优选是平坦的）; 第一导体和第二导体各自沿着支撑件延伸并且包括用于连接到电路的装置。 定位成接触液体混合物和第一导体的活性电极包括能够催化涉及化合物和（优选）电子介体的反应的酶的沉积物，其能够在酶催化的反应和第一个 导体。 定位参考电极以接触混合物和第二导体。 该系统包括适于提供代表电流的电信号的电路。 双电极条的制造，例如通过将酶，导电材料和介体的混合物作为单层丝网印刷到基底上。
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