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1. 发明申请

US20050043521A1 Phage antibodies 审中-公开
标题翻译：噬菌体抗体
公开(公告)号：US20050043521A1
公开(公告)日：2005-02-24
申请号：US10910547
申请日：2004-08-03
申请人： Leon Terstappen , Ton Logtenberg
发明人： Leon Terstappen , Ton Logtenberg
IPC分类号： C07K16/28 , C12P21/04 , C12Q1/70 , C07K16/46
CPC分类号： C07K16/2887 , C07K16/2809 , C07K2317/21 , C07K2317/622
摘要： Peripheral blood leucocytes incubated with a semi-synthetic phage antibody library and fluorochrome-labeled CD3 and CD20 antibodies were used to isolate human single chain Fv antibodies specific for subsets of blood leucocytes by flow cytometry. Isolated phage antibodies showed exclusive binding to the subpopulation used for selection or displayed additional binding to a restricted population of other cells in the mixture. At least two phage antibodies appeared to display hithereto unknown staining patterns of B lineage cells. This approach provides a subtractive procedure to rapidly obtain human antibodies against known and novel surface antigens in their native configuration, expressed on phenotypically defined subpopulations of cells. Importantly, this approach does not depend on immunization procedures or the necessity to repeatedly construct phage antibody libraries.
摘要翻译：使用半合成噬菌体抗体文库和荧光染料标记的CD3和CD20抗体孵育的外周血白细胞通过流式细胞术分离对血液白细胞亚群特异性的人单链Fv抗体。分离的噬菌体抗体显示与用于选择的亚群的独特结合，或显示与混合物中其他细胞的限制群体的额外结合。至少有两种噬菌体抗体似乎显示出对B系细胞的未知染色模式的影响。这种方法提供了一种消减方法，以快速获得针对已知和新型表型抗原的人类抗体，其在天然配置中，在表型定义的细胞亚群中表达。重要的是，这种方法不依赖于免疫程序或重复构建噬菌体抗体文库的必要性。

2. 发明授权

US06551843B1 Methods for enhancing binding interactions between members of specific binding pairs 失效
标题翻译：增强特异性结合对成员之间结合相互作用的方法
公开(公告)号：US06551843B1
公开(公告)日：2003-04-22
申请号：US09240939
申请日：1999-01-29
申请人： Galla Chandra Rao , Leon Terstappen , Paul Liberti
发明人： Galla Chandra Rao , Leon Terstappen , Paul Liberti
IPC分类号： G01N3353
CPC分类号： G01N33/54333
摘要： Disclosed is a method that improves the efficiency of reactions between specific binding pairs. By translating one member of the pair through a suspension of the second member, or across a surface to which the second member is immobilized, collision and binding frequencies are greatly increased. The resulting increased binding frequency allows for reduced incubation times and lower incubation temperatures. The enhanced collisions are advantageous for both molecular reactions, in which mixing is not always effective, and cellular reactions, in which the cells may be damaged by mixing.
摘要翻译：公开了一种提高特异性结合对之间的反应效率的方法。通过将该对中的一个构件通过第二构件的悬架或通过固定有第二构件的表面平移，大大增加了碰撞和装订频率。所产生的增加的结合频率允许减少的孵育时间和较低的温育温度。增强的碰撞对于分子反应是有利的，其中混合并不总是有效，以及细胞反应，其中细胞可能被混合而损伤。

3. 发明申请

US20070154960A1 Method for assessing disease states by profile analysis of isolated circulating endothelial cells 有权
标题翻译：通过分离的循环内皮细胞的轮廓分析来评估疾病状态的方法
公开(公告)号：US20070154960A1
公开(公告)日：2007-07-05
申请号：US11701763
申请日：2007-02-02
申请人： Mark Connelly , Gerald Doyle , Galla Rao , Leon Terstappen
发明人： Mark Connelly , Gerald Doyle , Galla Rao , Leon Terstappen
IPC分类号： G01N33/567
CPC分类号： G01N33/6893 , G01N33/5091 , G01N2800/32 , Y10T436/101666 , Y10T436/107497 , Y10T436/108331 , Y10T436/2525 , Y10T436/25375
摘要： Elevated number of Circulating Endothelial Cells (CEC) have been implicated in disease conditions associated with the formation or destruction of blood vessels such as acute coronary syndrome, thrombocytopenic purpura, sickle cell disease, sepsis, lupus, nephrotic syndromes, rejection of organ transplants, surgical trauma and cancer. This invention provides a method for assessing the levels of CEC which vary between different studies using a sensitive enrichment, imaging, and enumberation analysis. CD146 is one of the most specific endothelium-associated cell-surface antigens which can be used in image cytometry. CEC analysis provides an essential tool in prognostic/diagnostic evaluation in the clinic.
摘要翻译：循环内皮细胞数量增加已涉及与血管形成或破坏相关的疾病状况，如急性冠状动脉综合征，血小板减少性紫癜，镰状细胞病，败血症，狼疮，肾病综合征，器官移植排斥，外科手术创伤和癌症。本发明提供了一种用于评估使用敏感浓缩，成像和启发分析的不同研究之间变化的CEC水平的方法。 CD146是可用于图像细胞计数的最特异的内皮相关细胞表面抗原之一。 CEC分析为诊所的预后/诊断评估提供了重要的工具。

4. 发明申请

US20050181463A1 Analysis of circulating tumor cells, fragments, and debris 有权
标题翻译：分析循环肿瘤细胞，碎片和碎片
公开(公告)号：US20050181463A1
公开(公告)日：2005-08-18
申请号：US10780399
申请日：2004-02-17
申请人： Galla Rao , Christopher Larson , Madeline Repollet , Herman Rutner , Leon Terstappen , Shawn O'Hara , Steven Gross
发明人： Galla Rao , Christopher Larson , Madeline Repollet , Herman Rutner , Leon Terstappen , Shawn O'Hara , Steven Gross
IPC分类号： G01N33/53 , G01N33/543 , G01N33/567 , G01N33/574
CPC分类号： G01N33/574 , G01N33/54326 , Y10S436/813 , Y10T436/101666 , Y10T436/25125 , Y10T436/25375
摘要： The methods and reagents described in this invention are used to analyze circulating tumor cells, clusters, fragments, and debris. Analysis is performed with a number of platforms, including flow cytometry and the CellSpotter® fluorescent microscopy imaging system. Analyzing damaged cells has shown to be important. However, there are two sources of damage: in vivo and in vitro. Damage in vivo occurs by apoptosis, necrosis, or immune response. Damage in vitro occurs during sample acquisition, handling, transport, processing, or analysis. It is therefore desirable to confine, reduce, eliminate, or at least qualify in vitro damage to prevent it from interfering in analysis. Described herein are methods to diagnose, monitor, and screen disease based on circulating rare cells, including malignancy as determined by CTC, clusters, fragments, and debris. Also provided are kits for assaying biological specimens using these methods.
摘要翻译：本发明描述的方法和试剂用于分析循环肿瘤细胞，簇，碎片和碎片。使用许多平台进行分析，包括流式细胞仪和CellSpotter®荧光显微镜成像系统。分析损伤的细胞已被证明是重要的。然而，有两个损害来源：体内和体外。体内损伤是通过细胞凋亡，坏死或免疫应答发生的。在样品采集，处理，运输，加工或分析过程中发生体外损伤。因此，期望限制，减少，消除或至少限定体外损伤，以防止其干扰分析。本文描述了基于循环稀有细胞（包括由CTC，簇，碎片和碎片确定的恶性肿瘤）来诊断，监测和筛选疾病的方法。还提供了使用这些方法测定生物样品的试剂盒。

5. 发明申请

US20070117158A1 Methods and algorithms for cell enumeration in a low-cost cytometer 有权
标题翻译：低成本细胞计数器细胞计数的方法和算法
公开(公告)号：US20070117158A1
公开(公告)日：2007-05-24
申请号：US11434321
申请日：2006-05-12
申请人： Frank Coumans , Jan Greve , Frank Modica , Leon Terstappen , Arian Tibbe , John Verrant
发明人： Frank Coumans , Jan Greve , Frank Modica , Leon Terstappen , Arian Tibbe , John Verrant
IPC分类号： G06T7/20 , G01N33/567 , C12M1/34
CPC分类号： C12M41/36 , G01N15/1475
摘要： The enumeration of cells in fluids by flow cytometry is widely used across many disciplines such as assessment of leukocyte subsets in different bodily fluids or of bacterial contamination in environmental samples, food products and bodily fluids. For many applications the cost, size and complexity of the instruments prevents wider use, for example, CD4 analysis in HIV monitoring in resource-poor countries. The novel device, methods and algorithms disclosed herein largely overcome these limitations. Briefly, all cells in a biological sample are fluorescently labeled, but only the target cells are also magnetically labeled. In addition, non-magnetically labeled cells are imaged for viability in a modified slide configuration. The labeled sample, in a chamber or cuvet, is placed between two wedge-shaped magnets to selectively move the magnetically labeled cells to the observation surface of the cuvet. An LED illuminates the cells and a CCD camera captures the images of the fluorescent light emitted by the target cells. Image analysis performed with a novel algorithm provides a count of the cells on the surface that can be related to the target cell concentration of the original sample. The compact cytometer system provides a rugged, affordable and easy-to-use technique, which can be used in remote locations.
摘要翻译：通过流式细胞术对流体细胞的计数被广泛应用于许多学科，例如评估不同体液中的白细胞亚群或环境样品，食品和体液中的细菌污染。对于许多应用，仪器的成本，尺寸和复杂性阻止了更广泛的使用，例如资源贫乏国家的艾滋病毒监测中的CD 4分析。本文公开的新颖的装置，方法和算法在很大程度上克服了这些限制。简言之，生物样品中的所有细胞都被荧光标记，但只有靶细胞也被磁性标记。另外，非磁性标记的细胞在修改的载玻片配置中的存活力被成像。将标记的样品置于室或铜管中，放置在两个楔形磁体之间，以选择性地将磁性标记的细胞移动到铜管的观察表面。 LED照亮细胞，CCD相机捕获由靶细胞发射的荧光的图像。用新算法进行的图像分析提供了与原始样品的靶细胞浓度相关的表面上的细胞计数。紧凑型细胞仪系统提供坚固，经济实惠且易于使用的技术，可用于偏远地区。

6. 发明申请

US20060257847A1 Magnetic separation apparatus and methods 有权
标题翻译：磁分离装置及方法
公开(公告)号：US20060257847A1
公开(公告)日：2006-11-16
申请号：US11447562
申请日：2006-06-06
申请人： Tycho Scholtens , Leon Terstappen , Arjan Tibbe
发明人： Tycho Scholtens , Leon Terstappen , Arjan Tibbe
IPC分类号： C12Q1/00 , C12M1/34
CPC分类号： C12M33/00 , Y10T436/25 , Y10T436/25375
摘要： Apparatuses and methods for separating, immobilizing, and quantifying biological substances from within a fluid medium. Biological substances are observed by employing a vessel (6) having a chamber therein, the vessel comprising a transparent collection wall (5). A high internal gradient magnetic capture structure may be on the transparent collection wall (5), magnets (3) create an externally-applied force for transporting magnetically responsive material toward the transparent collection wall (5). V-shaped grooves on the inner surface of the viewing face of the chamber provide uniform. The invention is also useful in conducting quantitative analysis and sample preparation in conjunction with automated cell enumeration techniques.
摘要翻译：从流体介质中分离，固定和定量生物物质的装置和方法。通过使用其中具有室的容器（6）观察生物物质，所述容器包括透明收集壁（5）。高的内部梯度磁捕获结构可以在透明收集壁（5）上，磁体（3）产生用于将磁响应材料传送到透明收集壁（5）的外部施加的力。在腔室的观察面的内表面上的V形凹槽提供均匀的。本发明还可用于结合自动细胞计数技术进行定量分析和样品制备。

7. 发明申请

US20060024756A1 Methods and algorithms for cell enumeration in low-cost cytometer 有权
标题翻译：低成本细胞计数器细胞计数的方法和算法
公开(公告)号：US20060024756A1
公开(公告)日：2006-02-02
申请号：US10903798
申请日：2004-07-30
申请人： Arjan Tibbe , Jan Greve , Dhanesh Gohel , Erik Droog , Leon Terstappen
发明人： Arjan Tibbe , Jan Greve , Dhanesh Gohel , Erik Droog , Leon Terstappen
IPC分类号： G01N33/53
CPC分类号： G01N15/1475 , G01N15/1463 , G01N33/5094 , G01N33/54333 , G01N2015/008 , G01N2015/1486 , Y10T436/101666 , Y10T436/25375
摘要： The enumeration of cells in fluids by flow cytometry is widely used across many disciplines such as assessment of leukocyte subsets in different bodily fluids or of bacterial contamination in environmental samples, food products and bodily fluids. For many applications the cost, size and complexity of the instruments prevents wider use, for example, CD4 analysis in HIV monitoring in resource-poor countries. The novel device, methods and algorithms disclosed herein largely overcome these limitations. Briefly, all cells in a biological sample are fluorescently labeled, but only the target cells are also magnetically labeled. The labeled sample, in a chamber or cuvet, is placed between two wedge-shaped magnets to selectively move the magnetically labeled cells to the observation surface of the cuvet. An LED illuminates the cells and a CCD camera captures the images of the fluorescent light emitted by the target cells. Image analysis performed with a novel algorithm provides a count of the cells on the surface that can be related to the target cell concentration of the original sample. The compact cytometer system provides a rugged, affordable and easy-to-use technique, which can be used in remote locations.
摘要翻译：通过流式细胞术对流体细胞的计数被广泛应用于许多学科，例如评估不同体液中的白细胞亚群或环境样品，食品和体液中的细菌污染。对于许多应用，仪器的成本，尺寸和复杂性阻止了更广泛的使用，例如资源贫乏国家的艾滋病毒监测中的CD4分析。本文公开的新颖的装置，方法和算法在很大程度上克服了这些限制。简言之，生物样品中的所有细胞都被荧光标记，但只有靶细胞也被磁性标记。将标记的样品置于室或铜管中，放置在两个楔形磁体之间，以选择性地将磁性标记的细胞移动到铜管的观察表面。 LED照亮细胞，CCD相机捕获由靶细胞发射的荧光的图像。用新算法进行的图像分析提供了与原始样品的靶细胞浓度相关的表面上的细胞计数。紧凑型细胞仪系统提供坚固，经济实惠且易于使用的技术，可用于偏远地区。

8. 发明申请

US20070037173A1 Circulating tumor cells (CTC's): early assessment of time to progression, survival and response to therapy in metastatic cancer patients 审中-公开
标题翻译：循环肿瘤细胞（CTC）：转移性癌症患者的早期评估进展时间，存活和治疗反应
公开(公告)号：US20070037173A1
公开(公告)日：2007-02-15
申请号：US11202875
申请日：2005-08-12
申请人： Jeffrey Allard , Massimo Cristofanilli , Leon Terstappen
发明人： Jeffrey Allard , Massimo Cristofanilli , Leon Terstappen
IPC分类号： C12Q1/68 , G01N33/574 , G06F19/00
CPC分类号： G01N33/574 , G01N33/54326
摘要： A cancer test having prognostic utility in predicting time to disease progression, overall survival, and response to therapy in patients with MBC based upon the presence and number of CTC's. The Cell Spotter® System is used to enumerate CTC's in blood. The system immunomagnetically concentrates epithelial cells, fluorescently labels the cells and identifies and quantifies CTC's. The absolute number of CTC's detected in the peripheral blood tumor load is, in part, a factor in prediction of survival, time to progression, and response to therapy. The mean time to survival of patients depended upon a threshold number of 5 CTC's per 7.5 ml of blood. Detection of CTC's in metastatic cancer represents a novel prognostic factor in patients with metastatic cancers, suggests a biological role for the presence of tumor cells in the blood, and indicates that the detection of CTC's could be considered an appropriate surrogate marker for prospective therapeutic clinical trials.
摘要翻译：基于CTC的存在和数量，一种具有预测用于预测疾病进展时间，总体存活和对MBC患者的治疗反应的癌症测试。 Cell Spotter（R）系统用于列举血液中的四氯化碳。该系统免疫磁性浓缩上皮细胞，荧光标记细胞，并识别和量化CTC。在外周血肿瘤负荷中检测到的四氯化碳的绝对数量部分是预测生存期，进展时间和治疗反应的因素。患者生存的平均时间取决于每7.5毫升血液的5个CTC的阈值数。在转移性癌症中检测CTC代表转移癌患者的新型预后因素，表明血液中肿瘤细胞存在的生物学作用，并且表明CTC检测可被认为是前瞻性治疗性临床试验的适当替代标记。

9. 发明申请

US20060194192A1 Stabilization of cells and biological specimens for analysis 审中-公开
标题翻译：稳定细胞和生物标本进行分析
公开(公告)号：US20060194192A1
公开(公告)日：2006-08-31
申请号：US11413544
申请日：2006-04-28
申请人： Galla Rao , Melissa Herman , Herman Rutner , Leon Terstappen
发明人： Galla Rao , Melissa Herman , Herman Rutner , Leon Terstappen
IPC分类号： C12N5/00 , A01N1/02
CPC分类号： A01N1/02 , A01N1/0226 , A61K31/198 , A61K31/727
摘要： Compositions and methods for stabilizing rare cells in blood specimens, preserving the quality of blood specimens, and also serving as cell fixatives are disclosed which minimize losses of target cells (for example, circulating tumor cells) and formation of debris and aggregates from target cells, non-target cells and plasma components, thereby allowing more accurate analysis and classification of circulating tumor cells (CTC) and, ultimately, of tumor burdens in cancer patients. Stabilization of specimens is particularly desirable in protocols requiring rare cell enrichment from blood specimens drawn from cancer patients. Exposure of such specimens to potentially stressful conditions encountered, for example, in normal processing, mixing, shaking, delays due to transporting the blood, has been observed to not only diminish the number of CTC but also to generate debris and aggregates in the blood specimens that were found to interfere with accurate enumeration of target cells, if present. Stabilizers are necessary to discriminate between in vivo CTC disintegration and in vitro sample degredation.
摘要翻译：公开了用于稳定血液标本中稀有细胞，保持血液标本质量以及用作细胞固定剂的组合物和方法，其使靶细胞（例如循环肿瘤细胞）的损失最小化，并从靶细胞形成碎片和聚集体，非靶细胞和血浆成分，从而允许更准确地分析和分类循环肿瘤细胞（CTC），最终导致癌症患者的肿瘤负担。在从癌症患者抽取的血液标本中需要罕见细胞富集的方案中，标本的稳定是特别需要的。已经观察到这些样品暴露于例如在正常加工，混合，摇动，由于运送血液引起的延迟所遇到的潜在压力条件下，不仅减少了CTC的数量，而且在血样中产生碎片和聚集体被发现干扰靶细胞的准确计数（如果存在）。鉴别体内CTC分解和体外样品降解是必需的稳定剂。

10. 发明申请

US20060147901A1 Devices and methods to image objects by time delay integration 审中-公开
标题翻译：通过延时整合对物体进行成像的设备和方法
公开(公告)号：US20060147901A1
公开(公告)日：2006-07-06
申请号：US11344757
申请日：2006-02-01
申请人： Greve Jan , Federik Schreuder , Leon Terstappen , Arjan Tibbe
发明人： Greve Jan , Federik Schreuder , Leon Terstappen , Arjan Tibbe
IPC分类号： C12Q1/00 , G06K9/00
CPC分类号： G01N33/54326
摘要： Devices and methods for automated collection and image analysis are disclosed that enable identification or classification of microscopic objects aligned or deposited on surfaces. Such objects, e.g. detectably labeled rare target cells, are magnetically or non-magnetically immobilized and subjected to Time Delay Integration imaging (TDI). Incorporation of TDI technology into image cytometry analysis, like CellTracks®, makes it possible to image moving objects with very high sensitivity and signal-to-noise ratios. Implementation of TDI camera technology with dual excitation and multispectral imaging of enriched rare cells provides a rapid system for detection, enumeration, differentiation and characterization of imaged rare cells on the basis of size, morphology and immunophenotype.
摘要翻译：公开了用于自动收集和图像分析的装置和方法，其能够识别或分类对准或沉积在表面上的微观物体。这些物体，例如。可检测标记的罕见靶细胞是磁性或非磁性固定的并经受时间延迟积分成像（TDI）。将TDI技术结合到图像细胞分析中，像CellTracks（R），使得可以以非常高的灵敏度和信噪比来对移动物体进行成像。基于大小，形态和免疫表型，实现了具有双重激发和多光谱成像的富集稀有细胞的TDI相机技术，提供了快速检测，计数，分化和表征成像稀有细胞的系统。

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