公开(公告)号：WO1993010258A1

公开(公告)日：1993-05-27

申请号：PCT/US1992009873

申请日：1992-11-16

Applicant: BECKMAN INSTRUMENTS, INC.

Inventor： BECKMAN INSTRUMENTS, INC. ,  HSIEH, You-Zung ,  CHEN, Fu-Tai, A. ,  STERNBERG, James, C. ,  KLIEN, Gerald ,  LIU, Cheng-Ming

IPC: C12Q01/00

CPC classification number: C12Q1/007 ,  G01N27/44747

Abstract: Disclosed herein is a methodology for analyzing isoenzymes using capillary zone electrophoresis ('CZE') techniques. Briefly, an isoenzyme-containing sample and a substrate capable of being catalyzed by said isoenzyme into a reaction product are introduced into a capillary column comprising a buffer. Most preferably, the buffer contains the substrate prior to introduction of the sample into such substrate-buffer. CZE separation techniques are applied to the column such that the isoenzymes are separated from each other into discrete zones. The separation techniques are terminated such that product is rapidly generated by the catalytic conversion of substrate by the isoenzymes, and accumulated, within each discrete zone, followed by detection of product. Information regarding the relative distribution of the isoenzymes can be derived from the relative distribution of the product.

公开(公告)号：WO1990000252A1

公开(公告)日：1990-01-11

申请号：PCT/US1989002372

申请日：1989-05-31

Applicant: BECKMAN INSTRUMENTS, INC.

Inventor： BECKMAN INSTRUMENTS, INC. ,  STERNBERG, James, C.

IPC: G01N33/543

CPC classification number: G01N33/54306 ,  G01N33/581 ,  G01N33/582

Abstract: Specific binding assays for measuring the concentration of an analyte in a sample involve forming a complex comprising a solid support, the analyte or an analogue of the analyte, and an enzyme. The enzyme is used to cleave non-isotopic labels that are directly detectable from a substrate/label conjugate to provide a plurality of released labels in the free state, which are then separated from conjugate containing unreleased label. The released labels are detected for measuring the concentration of the analyte in the sample. Preferably the analyte or analogue is connected to the solid support by a cleavable bond, so that the enzyme can be separated from the solid support for use in the enzymatic cleaving of labels from the conjugate. This preferred version reduces the adverse effect of non-specific binding on the sensitivity and accuracy of the assay.

Abstract translation: 用于测量样品中分析物浓度的特异性结合测定包括形成包含固体支持物，分析物或分析物的类似物的复合物和酶。 该酶用于切割可从底物/标记缀合物直接检测的非同位素标记，以提供游离状态的多个释放的标记，然后将其与含有未释放标签的缀合物分离。 检测释放的标签以测量样品中分析物的浓度。 优选地，分析物或类似物通过可切割键连接到固体支持物，使得酶可以与固体支持物分离，用于从缀合物酶标记的酶切割。 该优选的形式减少非特异性结合对测定的灵敏度和准确性的不利影响。

公开(公告)号：WO1996006948A1

公开(公告)日：1996-03-07

申请号：PCT/US1995010226

申请日：1995-08-10

Applicant: BECKMAN INSTRUMENTS, INC.

Inventor： BECKMAN INSTRUMENTS, INC. ,  REDDY, M., Parameswara ,  STERNBERG, James, C.

IPC: C12Q01/68

CPC classification number: C12Q1/6825 ,  C12Q1/6804 ,  C12Q1/6813 ,  C12Q1/6816 ,  C12Q1/6834

Abstract: Double stranded nucleic acid duplexes serve as universal harvestable and cleavable link systems in a variety of different types of immnuoassays (e.g., sandwich, competitive, etc.). Depending upon the type of assay, at least one specific component involved in the assay system is attached to a first member of a pair of sequences forming a double stranded nucleic acid (i.e., two oligonucleotides comprising substantially complementary sequences). The assay is carried out in the presence of a support to which is attached an oligonucleotide which is the other member of the pair of sequences forming a double-stranded nucleic acid duplex under hybridization conditions. Upon the hybridization of the two complementary oligonucleotides to form a duplex, the component of the assay system to which the first member of the pair of oligonucleotides is attached may thereby be effectively removed from the solution phase and harvested onto the support. Oligonucleotides bound to a support are reusable in multiple successive assays. Moreover, any given support-bound oligonucleotide can be used in accordance with the present invention for the analysis of a variety of different analytes. In many cases, the assay system includes a label to facilitate quantifying the amount of analyte; in others, the amount of analyte may be determined without the use of any extraneous label.

Abstract translation: 双链核酸双链体在各种不同类型的免疫测定（例如，三明治，竞争性等）中用作通用可收获和可切割的连锁系统。 根据测定的类型，参与测定系统的至少一个特定组分连接到形成双链核酸的一对序列的第一个成员（即，包含基本上互补序列的两个寡核苷酸）。 所述测定在载体的存在下进行，其中连接寡核苷酸，其是在杂交条件下形成双链核酸双链体的一对序列的另一个成员。 当两个互补寡核苷酸杂交以形成双链体时，可以从溶液相中有效地从溶液相中除去与该对寡核苷酸的第一个成员所连接的测定系统的组分并收获到载体上。 结合支持物的寡核苷酸可在多次连续测定中重复使用。 此外，根据本发明，可以使用任何给定的支持结合寡核苷酸来分析各种不同的分析物。 在许多情况下，测定系统包括有助于量化分析物量的标签; 在其他方面，可以在不使用任何外来标签的情况下确定分析物的量。

公开(公告)号：WO1994017401A1

公开(公告)日：1994-08-04

申请号：PCT/US1994000762

申请日：1994-01-19

Applicant: BECKMAN INSTRUMENTS, INC.

Inventor： BECKMAN INSTRUMENTS, INC. ,  HERRICK, Steven, S. ,  STERNBERG, James, C.

IPC: G01N27/447

CPC classification number: G01N27/44795 ,  G01N27/44752

Abstract: The present invention provides processes for isoelectric focusing ("IEF") and associated detection, which incorporates a dynamic means of electroosmotic flow ("EOF") control during IEF and/or after IEF to effect solute mobilization. In accordance with the present invention, the EOF control during IEF and/or solute mobilization after IEF are accomplished by applying an external electric field, relative to an internal electric field, to modify the electroosmotic flow in the capillary. This can be done by disposing a conductive member at one or more locations outside and along the buffer column in the capillary. The conductive member may be statically charged or caused to conduct a current to create the external required electric field. The applied external electric field may be adjusted, relative to the external electric field, during IEF necessary to reduce or completely suppress EOF to prevent flow of the buffer. Upon the completion of IEF (irrespective of the method of reducing or removing EOF during IEF), the external electric field is adjusted, relative to the internal electric field, such that the buffer carrying the focused solutes are moved electroosmotically through the capillary past a detection point. The present invention is applicable to internally coated capillary which suppresses EOF even in the absence of the external electric field.

Abstract translation: 本发明提供等电聚焦（“IEF”）和相关检测的方法，其包括在IEF期间和/或在IEF之后的电渗流（“EOF”）控制的动态方式以进行溶质动员。 根据本发明，在IEF之间的IEF和/或溶质动员期间的EOF控制是通过相对于内部电场施加外部电场来实现的，以改变毛细管中的电渗流。 这可以通过在毛细管中的缓冲柱的外部和沿着一个或多个位置设置导电构件来实现。 导电构件可以被静电充电或导致电流以产生外部所需的电场。 在IEF期间，可以相对于外部电场调整所施加的外部电场，以减少或完全抑制EOF以防止缓冲器的流动。 在完成IEF（不管在IEF期间减少或去除EOF的方法）时，相对于内部电场调节外部电场，使得携带聚焦溶质的缓冲液通过毛细管电动移动通过检测 点。 本发明适用于即使没有外部电场也能抑制EOF的内部涂布毛细管。

公开(公告)号：WO1989003041A2

公开(公告)日：1989-04-06

申请号：PCT/US1988003368

申请日：1988-09-30

Applicant: BECKMAN INSTRUMENTS, INC.

Inventor： BECKMAN INSTRUMENTS, INC. ,  OH, Chan, S. ,  STERNBERG, James, C.

IPC: G01N33/531

CPC classification number: G01N33/54353 ,  G01N33/531 ,  G01N33/532 ,  G01N33/539 ,  G01N33/54313 ,  G01N33/94 ,  Y10S435/964 ,  Y10S436/819 ,  Y10S530/81 ,  Y10S530/812 ,  Y10S530/816

Abstract: A tridentate conjugate has three chemical moieties, or tridentate members, attached through an appropriate spacer moiety. At least two of the tridentate members are relatively small molecules, usually less than about 7,000 Daltons in size. The particular appropriate spacer moiety selected for a tridentate imparts certain steric properties to the tridentate conjugate. In one embodiment, the binding of a macromolecular specific binding partner to one of the tridentate members sterically inhibits the binding of a different macromolecule to another tridentate member. In another embodiment, the binding of a first tridentate member to a macromolecule restricts the subsequent binding of a second tridentate member to a proximate location on the same macromolecule.

Abstract translation: 具有三个化学部分或三齿元素的三齿共轭物通过适当的间隔半部连接。 至少两个三齿元素是相对小的分子，其大小通常小于7000道尔顿。 为三齿选择适当的半粒径赋予三齿偶联物一定的空间性质。 在一个实施方案中，将特定大分子结合配偶体与三齿元件之一结合，从空间上防止不同大分子与另一三齿元件的结合。 在另一个实施方案中，第一三齿元素与大分子的结合限制了第二三齿元素随后与同一分子上的附近位置的结合。

公开(公告)号：EP0632890A1

公开(公告)日：1995-01-11

申请号：EP94907281.0

申请日：1994-01-19

Applicant: BECKMAN INSTRUMENTS, INC.

Inventor： HERRICK, Steven, S. ,  STERNBERG, James, C.

IPC: B01D57 ,  G01N27

CPC classification number: G01N27/44795 ,  G01N27/44752

Abstract: The present invention provides processes for isoelectric focusing ('IEF') and associated detection, which incorporates a dynamic means of electroosmotic flow ('EOF') control during IEF and/or after IEF to effect solute mobilization. In accordance with the present invention, the EOF control during IEF and/or solute mobilization after IEF are accomplished by applying an external electric field, relative to an internal electric field, to modify the electroosmotic flow in the capillary. This can be done by disposing a conductive member at one or more locations outside and along the buffer column in the capillary. The conductive member may be statically charged or caused to conduct a current to create the external required electric field. The applied external electric field may be adjusted, relative to the external electric field, during IEF necessary to reduce or completely suppress EOF to prevent flow of the buffer. Upon the completion of IEF (irrespective of the method of reducing or removing EOF during IEF), the external electric field is adjusted, relative to the internal electric field, such that the buffer carrying the focused solutes are moved electroosmotically through the capillary past a detection point. The present invention is applicable to internally coated capillary which suppresses EOF even in the absence of the external electric field.

公开(公告)号：EP0383860A1

公开(公告)日：1990-08-29

申请号：EP89907479.0

申请日：1989-05-31

Applicant: BECKMAN INSTRUMENTS, INC.

Inventor： STERNBERG, James, C.

IPC: G01N33

CPC classification number: G01N33/54306 ,  G01N33/581 ,  G01N33/582

Abstract: Des méthodes de titrages par liaison spécifique, permettant de mesurer la concentration d'un analyte dans un échantillon, consistent à former un complexe comprenant un support solide, l'analyte ou un analogue de l'analyte, et une enzyme. On utilise l'enzyme pour cliver des marques non isotropiques directement détectables à partir d'un conjugué substrat/marque, afin de produire une pluralité de marques libérées dans l'état libre, que l'on sépare ensuite du conjugué contenant la marque non libérée. On détecte les marques libérées pour mesurer la concentration de l'analyte dans l'échantillon. L'analyte ou analogue est de préférence fixé au support solide par une liaison clivable, de sorte que l'on peut séparer l'enzyme du support solide pour l'utiliser dans le clivage enzymatique de marques à partir du conjugué. Cette version préférée réduit l'effet désavantageux d'une liaison non spécifique sur la sensibilité et la précision du titrage.

公开(公告)号：EP0779934A1

公开(公告)日：1997-06-25

申请号：EP95928819.0

申请日：1995-08-10

Applicant: BECKMAN INSTRUMENTS, INC.

Inventor： REDDY, M., Parameswara ,  STERNBERG, James, C.

IPC: G01N33 ,  C12N15 ,  C12Q1

CPC classification number: C12Q1/6825 ,  C12Q1/6804 ,  C12Q1/6813 ,  C12Q1/6816 ,  C12Q1/6834

Abstract: Double stranded nucleic acid duplexes serve as universal harvestable and cleavable link systems in a variety of different types of immnuoassays (e.g., sandwich, competitive, etc.). Depending upon the type of assay, at least one specific component involved in the assay system is attached to a first member of a pair of sequences forming a double stranded nucleic acid (i.e., two oligonucleotides comprising substantially complementary sequences). The assay is carried out in the presence of a support to which is attached an oligonucleotide which is the other member of the pair of sequences forming a double-stranded nucleic acid duplex under hybridization conditions. Upon the hybridization of the two complementary oligonucleotides to form a duplex, the component of the assay system to which the first member of the pair of oligonucleotides is attached may thereby be effectively removed from the solution phase and harvested onto the support. Oligonucleotides bound to a support are reusable in multiple successive assays. Moreover, any given support-bound oligonucleotide can be used in accordance with the present invention for the analysis of a variety of different analytes. In many cases, the assay system includes a label to facilitate quantifying the amount of analyte; in others, the amount of analyte may be determined without the use of any extraneous label.

公开(公告)号：EP0572604A1

公开(公告)日：1993-12-08

申请号：EP92925261.0

申请日：1992-11-16

Applicant: BECKMAN INSTRUMENTS, INC.

Inventor： HSIEH, You-Zung ,  CHEN, Fu-Tai, A. ,  STERNBERG, James, C. ,  KLIEN, Gerald ,  LIU, Cheng-Ming

IPC: C12Q1 ,  G01N27

CPC classification number: C12Q1/007 ,  G01N27/44747

Abstract: Procédé d'analyse d'isoenzymes utilisant des techniques d'électrophorèse à zone capillaire ("CZE"). Un échantillon contenant une isoenzyme et un substrat pouvant être catalysé par ladite isoenzyme en un produit de réaction sont introduits dans une colonne capillaire comprenant un tampon. Idéalement, le tampon contient le substrat avant l'introduction de l'échantillon. On applique à la colonne des techniques de séparation CZE, afin de séparer les isoenzymes en des zones discrètes. A la fin de la séparation, le produit est rapidement généré par la conversion catalytique du substrat par les isoenzymes, et accumulé dans chaque zone discrète. Le produit est ensuite détecté. On peut dériver les informations sur la répartition relative des isoenzymes à partir de la distribution relative du produit.

公开(公告)号：EP0572604B1

公开(公告)日：1997-05-07

申请号：EP92925261.7

申请日：1992-11-16

Applicant: BECKMAN INSTRUMENTS, INC.

Inventor： HSIEH, You-Zung ,  CHEN, Fu-Tai, A. ,  STERNBERG, James, C. ,  KLIEN, Gerald ,  LIU, Cheng-Ming

IPC: C12Q1/00 ,  G01N27/447

CPC classification number: C12Q1/007 ,  G01N27/44747

Abstract: Disclosed herein is a methodology for analyzing isoenzymes using capillary zone electrophoresis ('CZE') techniques. Briefly, an isoenzyme-containing sample and a substrate capable of being catalyzed by said isoenzyme into a reaction product are introduced into a capillary column comprising a buffer. Most preferably, the buffer contains the substrate prior to introduction of the sample into such substrate-buffer. CZE separation techniques are applied to the column such that the isoenzymes are separated from each other into discrete zones. The separation techniques are terminated such that product is rapidly generated by the catalytic conversion of substrate by the isoenzymes, and accumulated, within each discrete zone, followed by detection of product. Information regarding the relative distribution of the isoenzymes can be derived from the relative distribution of the product.