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1. 发明授权

US08323903B2 Antibody complexes and methods for immunolabeling 有权
标题翻译：抗体复合物和免疫标记方法
公开(公告)号：US08323903B2
公开(公告)日：2012-12-04
申请号：US10118204
申请日：2002-04-05
申请人： Robert A. Archer , Joseph M. Beechem , David C. Hagen , Richard P. Haugland , Rosaria P. Haugland
发明人： Robert A. Archer , Joseph M. Beechem , David C. Hagen , Richard P. Haugland , Rosaria P. Haugland
IPC分类号： G01N33/53
CPC分类号： B82Y30/00 , B82Y5/00 , B82Y10/00 , G01N33/53 , G01N33/58 , G01N33/6857
摘要： The present invention provides novel immunolabeling complexes and certain components of such complexes, as well as methods of preparing and using such complexes, and kits for use in preparing labeling proteins and for immunolabeling. The pre-formed immunolabeling complexes of the invention comprise both a target-binding antibody and a labeling protein that contains covalently attached labels, where the labeling protein binds selectively and with high affinity to a selected region of the target-binding antibody. Novel labeling proteins of the invention include non-antibody peptides and proteins, such as a complex of protein G and a labeled albumin, and monovalent antibody fragments, such as labeled Fab fragments of an anti-Fc antibody. In methods of the invention, the preformed immunolabeling complexes are added to the sample alone or in combination, for purposes of labeling and optionally detecting the target of interest.
摘要翻译：本发明提供了新的免疫标记复合物和这些复合物的某些成分，以及制备和使用这些复合物的方法，以及用于制备标记蛋白和免疫标记的试剂盒。本发明的预先形成的免疫粘附复合物包含靶结合抗体和含有共价连接的标记的标记蛋白，其中所述标记蛋白选择性地结合并且与靶结合抗体的选定区域具有高亲和力。本发明的新型标记蛋白包括非抗体肽和蛋白质，例如蛋白G和标记的白蛋白的复合物，以及单价抗体片段，例如抗Fc抗体的标记的Fab片段。在本发明的方法中，为了标记和任选地检测目的靶标，将预先形成的免疫标记复合物单独或组合加入到样品中。

2. 发明申请

US20110105362A1 CYANINE COMPOUNDS AND THEIR APPLICATION AS QUENCHING COMPOUNDS 审中-公开
标题翻译：氰胺化合物及其作为化学化合物的应用
公开(公告)号：US20110105362A1
公开(公告)日：2011-05-05
申请号：US13005002
申请日：2011-01-12
申请人： Rosaria P. Haugland , Ching-Ying Cheung , Stephen Yue
发明人： Rosaria P. Haugland , Ching-Ying Cheung , Stephen Yue
IPC分类号： C40B40/04 , C12N9/96 , C12N7/00 , C12N1/00 , C07K2/00 , C07K14/00 , C07K16/00 , C07H21/00 , C07H19/00 , C07D209/56 , C07D209/18
CPC分类号： C12Q1/6876 , C07D209/12 , C07D209/14 , C07D209/24 , C07D401/04 , C09B23/12
摘要： The present invention provides methods and non-fluorescent carbocyanine quencher compounds having the general formula: Wherein the A moiety is a substituted pyridinium, unsubstituted pyridinium, substituted quinolinium, unsubstituted quinolinium, substituted benzazolium, unsubstituted benzazolium, substituted indolinium, or substituted indolinium. The invention further provides luminescent donor molecule-quencher pairs and luminescent donor molecule-quencher-luminescent acceptor molecule conjugates wherein the quencher is a cyanine compound of the present invention. The energy transfer pairs are used to detect an analyte of interest in a sample.
摘要翻译：本发明提供了具有以下通式的方法和非荧光碳花青猝灭剂：其中A部分是取代的吡啶鎓，未取代的吡啶鎓，取代的喹啉鎓，未取代的喹啉鎓，取代的苯并唑鎓，未取代的苯并咪唑鎓，取代的吲哚鎓或取代的吲哚啉鎓。本发明还提供了发光供体分子猝灭剂对和发光供体分子 - 猝灭剂 - 发光受体分子缀合物，其中猝灭剂是本发明的花青化合物。能量转移对用于检测样品中感兴趣的分析物。

3. 发明申请

US20130072666A1 Antibody Complexes and Methods for Immunolabeling 有权
标题翻译：抗体复合物和免疫标记方法
公开(公告)号：US20130072666A1
公开(公告)日：2013-03-21
申请号：US13610009
申请日：2012-09-11
申请人： Robert M. Archer , Joseph M. Beechem , David C. Hagen , Richard P. Haugland , Rosaria P. Haugland
发明人： Robert M. Archer , Joseph M. Beechem , David C. Hagen , Richard P. Haugland , Rosaria P. Haugland
IPC分类号： C07K1/113 , C07K16/00 , C12P21/06
CPC分类号： B82Y5/00 , B82Y10/00 , G01N33/53
摘要： Provided are labeling reagents and methods for labeling primary antibodies and for detecting a target in a sample using an immuno-labeled complex that comprises a target-binding antibody and one or more labeling reagents. The labeling reagents comprise monovalent antibody fragments or non-antibody monomeric proteins whereby the labeling proteins have affinity for a specific region of the target-binding antibody and are covalently attached to a label. Discrete subsets of labeling reagent and immune-labeled complexes are provided that facilitate the simultaneous detection of multiple targets in a sample-complexes are distinguished by i) a ratio of label to labeling reagent, or ii) a physical property of said label, or iii) a ratio of labeling reagent to said target-binding antibody, or iv) by said target-binding antibody. This is particularly useful for fluorophore labels that can be attached to labeling reagents and subsequently immuno-labeled complexes in ratios for the detection of multiple targets.
摘要翻译：提供标记试剂和用于标记一抗的标记试剂和用于使用包含靶结合抗体和一种或多种标记试剂的免疫标记复合物检测样品中的靶标的方法。标记试剂包括单价抗体片段或非抗体单体蛋白质，由此标记的蛋白质对靶标结合抗体的特定区域具有亲和力，并且共价连接到标记物上。提供标记试剂和免疫标记复合物的离散子集，其有利于同时检测样品中多个靶标的复合物，其特征在于i）标记与标记试剂的比例，或ii）所述标记的物理性质，或iii ）标记试剂与所述靶结合抗体的比例，或iv）所述靶结合抗体。这对于可以连接到标记试剂和随后的用于检测多个靶的比例的免疫标记的复合物的荧光团标记物特别有用。

4. 发明申请

US20090124511A1 ANTIBODY COMPLEXES AND METHODS FOR IMMUNOLABELING 有权
标题翻译：抗体复合物和免疫标记方法
公开(公告)号：US20090124511A1
公开(公告)日：2009-05-14
申请号：US12047131
申请日：2008-03-12
申请人： Robert M. Archer , Joseph M. Beechem , David C. Hagen , Richard P. Haugland , Rosaria P. Haugland
发明人： Robert M. Archer , Joseph M. Beechem , David C. Hagen , Richard P. Haugland , Rosaria P. Haugland
IPC分类号： C40B30/04 , C07K1/13 , C40B50/00 , C12N9/00 , C12N9/08 , C12N9/02 , C40B50/08 , C40B40/10 , C07K16/00 , C12P21/06 , C12N9/16 , C12N9/24 , G01N33/567 , G01N33/543
CPC分类号： B82Y5/00 , B82Y10/00 , G01N33/53
摘要： The present invention provides labeling reagents and methods for labeling primary antibodies and for detecting a target in a sample using an immuno-labeled complex that comprises a target-binding antibody and one or more labeling reagents. The labeling reagents comprise monovalent antibody fragments or non-antibody monomeric proteins whereby the labeling proteins have affinity for a specific region of the target-binding antibody and are covalently attached to a label. Typically, the labeling reagent is an anti-Fc Fab or Fab′ fragment that was generated by immunizing a goat or rabbit with the Fc fragment of an antibody. The present invention provides for discrete subsets of labeling reagent and immuno-labeled complexes that facilitate the simultaneous detection of multiple targets in a sample wherein the immuno-labeled complexes are distinguished by i) a ratio of label to labeling reagent, or ii) a physical property of said label, or iii) a ratio of labeling reagent to said target-binding antibody, or iv) by said target-binding antibody. This is particularly useful for fluorophore labels that can be attached to labeling reagents and subsequently immuno-labeled complexes in ratios for the detection of multiple targets.
摘要翻译：本发明提供标记试剂和用于标记一抗的标记试剂和使用包含靶结合抗体和一种或多种标记试剂的免疫标记复合物检测样品中靶标的方法。标记试剂包括单价抗体片段或非抗体单体蛋白质，由此标记的蛋白质对靶标结合抗体的特定区域具有亲和力，并且共价连接到标记物上。通常，标记试剂是通过用抗体的Fc片段免疫山羊或兔而产生的抗Fc Fab或Fab'片段。本发明提供标记试剂和免疫标记的复合物的离散子集，其有助于同时检测样品中的多个靶标，其中通过i）标记物与标记试剂的比率来区分免疫标记的复合物，或ii）物理或iii）标记试剂与所述靶结合抗体的比例，或iv）所述靶结合抗体。这对于可以连接到标记试剂和随后的用于检测多个靶的比例的免疫标记的复合物的荧光团标记物特别有用。

5. 发明授权

US5723218A Dipyrrometheneboron difluoride labeled flourescent microparticles 失效
标题翻译：二吡咯甲烷二氟化物标记的荧光微粒
公开(公告)号：US5723218A
公开(公告)日：1998-03-03
申请号：US484151
申请日：1995-06-07
申请人： Richard P. Haugland , Rosaria P. Haugland , John Michael Brinkley , Hee Chol Kang , Michael Kuhn , K. Sam Wells , Yu Zhong Zhang
发明人： Richard P. Haugland , Rosaria P. Haugland , John Michael Brinkley , Hee Chol Kang , Michael Kuhn , K. Sam Wells , Yu Zhong Zhang
IPC分类号： C09K11/06 , G01N33/533 , B32B27/18
CPC分类号： G01N33/533 , C09K11/06 , Y10T428/2982 , Y10T428/2991 , Y10T428/2998
摘要： The invention is a novel fluorescently labeled microparticle, where the microparticle internally incorporates at least one dipyrrometheneboron difluoride dye. Appropriate selection of substituents results in dipyrrometheneboron difluoride derivatives that, when incorporated into polymer microparticles, give the desired excitation and emission wavelengths. The spectral characteristics of the labeling dyes in liquid are not greatly changed when the dye is incorporated into the particles, and the spectral excitation and emission wavelengths are compatible with commonly used filter sets. Other embodiments of the fluorescent microparticles include additional dyes and/or bioreactive substances.
摘要翻译：本发明是一种新型荧光标记微粒，其中微粒内部掺入至少一种二吡咯亚甲基二氟化物染料。取代基的适当选择导致二吡咯亚甲基二氟化物衍生物，当掺入聚合物微粒中时，得到期望的激发和发射波长。当染料掺入颗粒中时，液体中标记染料的光谱特性没有大的变化，光谱激发和发射波长与通常使用的滤光片组兼容。荧光微粒的其它实施方案包括另外的染料和/或生物反应性物质。

6. 发明授权

US5362628A Fluorescent haloalkyl derivatives of reporter molecules well retained in cells 失效
标题翻译：报道分子的荧光卤代烷基衍生物保留在细胞中
公开(公告)号：US5362628A
公开(公告)日：1994-11-08
申请号：US26633
申请日：1993-03-05
申请人： Richard P. Haugland , Yu-Zhong Zhang , Ram Sabnis , Nels A. Olson , John J. Naleway , Rosaria P. Haugland
发明人： Richard P. Haugland , Yu-Zhong Zhang , Ram Sabnis , Nels A. Olson , John J. Naleway , Rosaria P. Haugland
IPC分类号： C12Q1/04 , C12Q1/34 , G01N33/535 , C12Q1/00 , A61K37/02 , C07H15/24 , G01N33/40
CPC分类号： C12Q1/04 , C12Q1/34 , G01N33/535 , C12Q2334/00 , C12Q2334/20 , C12Q2334/40 , C12Q2337/20 , G01N2333/924 , Y10S530/802
摘要： The subject invention provides a method for analyzing the metabolic activity in cells by improving the retention of a detectable reporter molecule only in intact cells where a particular enzyme is present. In particular, improved retention results from a two part process involving conjugation of haloalkyl-substituted derivatives of a reporter molecule with intracellular cysteine-containing peptides while unblocking the reporter molecule. The method for analyzing metabolic activity of cells involves the use of a substrate having the formXR-REPORTER-BLOCKwherein -BLOCK is a group selected to be removable by action of a specific analyte, to give REPORTER spectral properties different from those of the substrate,-REPORTER- is a molecule that, when no longer bound to BLOCK by a BLOCK-REPORTER bond, has spectral properities different from those of the substrate, andXR-- is a haloalkyl moiety that can covalently react with an intracellular thiol (Z--S--H) to form a thioether conjugate (Z--S--R--).After the substrate enters the cells, the analyte removes BLOCK to make REPORTER detectable by the change in spectral properties, and the haloalkyl XR reacts with the intracellular thiol to form the thioether conjugate inside the cells, which is well-retained in the cells.
摘要翻译：本发明提供了通过仅在存在特定酶的完整细胞中改善可检测报道分子的保留来分析细胞中的代谢活性的方法。特别地，涉及将报道分子的卤代烷基取代的衍生物与含细胞内半胱氨酸的肽结合的两部分方法的改进的保留结果，同时解除报道分子的阻断。用于分析细胞代谢活性的方法涉及使用具有XR-REPORTER-BLOCK形式的底物，其中-BLOCK是通过特定分析物的作用被选择为可去除的基团，以产生与底物的不同的REPORTER光谱性质 -REPORTER-是一个分子，当不再通过BLOCK-REPORTER键与BLOCK结合时，具有与底物不同的光谱性质，XR-是可以与胞内硫醇（ZSH）共价反应的卤代烷基部分，以形成硫醚缀合物（ZSR-）。在底物进入细胞后，分析物除去BLOCK以通过光谱性质的变化使REPORTER可检测到，并且卤代烷基XR与细胞内硫醇反应以在细胞内形成硫醚缀合物，其在细胞中良好保留。

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