公开(公告)号：US5976822A

公开(公告)日：1999-11-02

申请号：US915414

申请日：1997-08-20

申请人： Eileen Landrum ,  Adry Galiounghi ,  Nancy Garcia ,  Ursino Del Valle ,  Frank J. Lucas

发明人： Eileen Landrum ,  Adry Galiounghi ,  Nancy Garcia ,  Ursino Del Valle ,  Frank J. Lucas

IPC分类号： C12Q1/26 ,  C12Q1/34 ,  C12Q1/37 ,  G01N33/574 ,  G01N33/58 ,  C12Q1/00

CPC分类号： C12Q1/26 ,  C12Q1/34 ,  C12Q1/37 ,  G01N33/574 ,  G01N33/582 ,  G01N2333/95 ,  G01N2333/96413 ,  G01N2333/96466 ,  G01N2333/96469 ,  G01N2800/52 ,  Y10S435/968

摘要： The ability to determine the stage or pathway of cysteine proteases in a single cell assay has long been desired as a material event in apoptosis. The present invention relates to a method and assay reagents for determining enzyme activity and relating said activity to the apoptotic pathway. In addition, the method find utility in distinguishing apoptotic activity from necrotic activity.

摘要翻译： 在单细胞测定中确定半胱氨酸蛋白酶的阶段或途径的能力早已被期望作为凋亡中的物质事件。 本发明涉及用于测定酶活性并将所述活性与凋亡途径相关联的方法和测定试剂。 此外，该方法可用于区分凋亡活性与坏死活性。

公开(公告)号：US5968831A

公开(公告)日：1999-10-19

申请号：US739362

申请日：1996-10-29

申请人： Ravindra S. Shukla ,  Harold R. Crews ,  Adry Galiounghi ,  Eileen Landrum ,  Frank J. Lucas

发明人： Ravindra S. Shukla ,  Harold R. Crews ,  Adry Galiounghi ,  Eileen Landrum ,  Frank J. Lucas

IPC分类号： G01N33/96 ,  A01N1/02 ,  C12N5/06 ,  C12N5/08 ,  C12N9/14 ,  G01N33/50 ,  G01N31/00 ,  C12Q1/00 ,  C12Q1/34

CPC分类号： G01N33/5005 ,  Y10T436/10

摘要： The invention provides a cell control product for use with a cytoenzymology assay to confirm enzymatic activity in devices employing electronic and/or optical means. The cell control product comprises a lyophilized mammalian cell which is capable of being rehydrated in water to exhibit cellular structure and cellular enzymatic activity so that said lyophilized cells can function effectively as a cytoenzymology cell control in an enzymatic analysis. The cellular structure of the cell control is capable of being authenticated by light scatter or microscopy analysis and the cellular enzymatic activity is capable of being authenticated by fluorescence analysis. The cell control exhibits a real time stability when stored at between 2 and 8.degree. C. for at least a two month period of time. Preferably the mammalian cell is an abnormal cell selected from Molt 4, CCRF-CEM and HL-60 cell lines.

摘要翻译： 本发明提供了一种细胞控制产品，用于细胞酶学测定以确认采用电子和/或光学方法的装置中的酶活性。 细胞对照产物包括冻干的哺乳动物细胞，其能够在水中再水化以显示细胞结构和细胞酶活性，使得所述冻干细胞可以有效地起到酶分析中的细胞酶学细胞控制的作用。 细胞控制的细胞结构能够通过光散射或显微镜分析进行鉴定，并且细胞酶活性能够通过荧光分析进行鉴定。 当在2℃和8℃之间储存至少两个月的时间时，细胞控制呈现实时稳定性。 优选地，哺乳动物细胞是选自Molt4，CCRF-CEM和HL-60细胞系的异常细胞。

公开(公告)号：US06692968B2

公开(公告)日：2004-02-17

申请号：US09957234

申请日：2001-09-20

申请人： Alexander Burshteyn ,  John W. Joubran ,  Nazle Kuylen ,  Frank J. Lucas ,  Carlos L. Aparicio ,  Michael L. Bell ,  Ravinder Gupta ,  Maria Elena Insausti ,  Jack D. McNeal ,  Paul W. Price ,  Sandra Socarras

发明人： Alexander Burshteyn ,  John W. Joubran ,  Nazle Kuylen ,  Frank J. Lucas ,  Carlos L. Aparicio ,  Michael L. Bell ,  Ravinder Gupta ,  Maria Elena Insausti ,  Jack D. McNeal ,  Paul W. Price ,  Sandra Socarras

IPC分类号： G01N134

CPC分类号： G01N1/4077 ,  B01D61/147 ,  B01D61/18 ,  B01D61/20 ,  B01D63/02 ,  Y10T436/25375 ,  Y10T436/255

摘要： A method for utilizing a filtration device for removing interferants from a test sample containing a mixture of a composition of interest and interferants in an automated apparatus is disclosed. The filtration device includes a microporous hollow fiber membrane having a plurality of pores sized to retain the composition of interest while allowing smaller diameter interferants to pass through the membrane.

摘要翻译： 公开了一种在自动化设备中利用过滤装置从含有感兴趣组合物和干扰物的混合物的测试样品中除去干扰物的方法。 过滤装置包括具有多个孔的微孔中空纤维膜，其尺寸适于保持感兴趣的组合物，同时允许较小直径的干燥剂通过膜。

公开(公告)号：US5698411A

公开(公告)日：1997-12-16

申请号：US444051

申请日：1995-05-18

申请人： Frank J. Lucas ,  Gerald E. Jaffe ,  Steven E. Bott ,  James H. Carter

发明人： Frank J. Lucas ,  Gerald E. Jaffe ,  Steven E. Bott ,  James H. Carter

IPC分类号： C12Q1/26 ,  C12Q1/34 ,  C12Q1/37 ,  G01N33/574 ,  G01N33/58 ,  C12Q1/02 ,  C12Q1/00

CPC分类号： G01N33/574 ,  C12Q1/26 ,  C12Q1/34 ,  C12Q1/37 ,  G01N33/582 ,  G01N2333/95 ,  G01N2333/96413 ,  G01N2333/96466 ,  G01N2333/96469 ,  G01N2800/52

摘要： A method for assaying the activity of an enzyme inside a metabolically active whole cell is disclosed. The assay compound includes a leaving group and an indicator group. The leaving group is selected from the group comprising amino acids, peptides, saccharides, sulfates, phosphates, esters, phosphate esters, nucleotides, polynucleotides, nucleic acids, pyrimidines, purines, nucleosides, lipids and mixtures thereof. The indicator group is selected from compounds which have a first state when joined to the leaving group, and a second state when the leaving group is cleaved from the indicator group by the enzyme. Preferably, the indicator compounds are rhodamine 110, rhodol, and fluorescein and analogs of these compounds. A method of synthesizing the compound is also disclosed.

摘要翻译： 公开了一种用于测定代谢活性全细胞内酶的活性的方法。 测定化合物包括离去基团和指示剂组。 离去基团选自包括氨基酸，肽，糖，硫酸盐，磷酸盐，酯，磷酸酯，核苷酸，多核苷酸，核酸，嘧啶，嘌呤，核苷，脂质及其混合物的组。 指示基团选自与离去基团连接时具有第一状态的化合物，当离去基团被酶从指示剂基团切割时的第二种状态。 优选地，指示剂化合物是这些化合物的罗丹明110，rhodol和荧光素以及类似物。 还公开了合成该化合物的方法。

公开(公告)号：US07354773B2

公开(公告)日：2008-04-08

申请号：US10437695

申请日：2003-05-14

申请人： Oilda Rubio ,  Carlos Aparicio ,  John A. Maples ,  Julie Wilkinson ,  Cecilia Smith ,  Frank J. Lucas

发明人： Oilda Rubio ,  Carlos Aparicio ,  John A. Maples ,  Julie Wilkinson ,  Cecilia Smith ,  Frank J. Lucas

IPC分类号： G01N1/18

CPC分类号： G01N1/30 ,  G01N1/405 ,  G01N15/147 ,  G01N33/5005 ,  G01N33/536 ,  Y10S435/971 ,  Y10T436/107497 ,  Y10T436/111666 ,  Y10T436/113332 ,  Y10T436/25375

摘要： A method and apparatus for preparing biological cell samples for intracellular analysis. The invention is based upon the recognition that many of the steps of the conventional methods for such sample preparation can be eliminated, leading to a process that readily lends itself to automation and the advantages associated therewith. The method of the invention comprises the steps of (a) cell-fixation, (b) permeabilization and (c) staining (or labeling) of intracellular molecules of interest by probes that are readily detectable by flow cytometric techniques, all without any intervening cell-washing (and re-suspension) steps. Rather, the single cell-washing step is effected after these three steps have been carried out. Preferably, the washing step is carried out by passing the fixed, permeabilized and stained cell sample through a semi-permeable membrane that serves to filter out (by transmission) interferants to waste while retaining the cells of interest.

摘要翻译： 一种用于制备用于细胞内分析的生物细胞样品的方法和装置。 本发明基于这样的认识，即可以消除用于这种样品制备的常规方法的许多步骤，导致易于适应自动化的过程以及与其相关的优点。 本发明的方法包括以下步骤：（a）细胞固定，（b）透化和（c）通过流式细胞术技术容易检测的探针染色（或标记）感兴趣的细胞内分子，所有这些都不具有任何中间细胞 洗涤（和重新暂停）步骤。 相反，在进行这三个步骤之后实现单细胞洗涤步骤。 优选地，洗涤步骤通过使固定的，透化的和染色的细胞样品通过半透膜进行，半透膜用于将（通过传输）干燥剂过滤掉，同时保留感兴趣的细胞。

公开(公告)号：US5849513A

公开(公告)日：1998-12-15

申请号：US904400

申请日：1997-07-31

申请人： Gerald E. Jaffe ,  Frank J. Lucas ,  James H. Carter

发明人： Gerald E. Jaffe ,  Frank J. Lucas ,  James H. Carter

IPC分类号： C12Q1/02 ,  C12Q1/34 ,  G01N33/574 ,  C12Q1/00

CPC分类号： C12Q1/34 ,  C12Q1/02 ,  G01N33/574 ,  C12Q2334/40 ,  C12Q2337/40

摘要： An assay compound or a salt thereof for assaying the activity of an enzyme inside a metabolically active whole cell is disclosed. The assay compound includes a leaving group and an indicator group. The leaving group is selected from the group comprising amino acids, peptides, saccharides, sulfates, phosphates, esters, phosphate esters, nucleotides, polynucleotides, nucleic acids, pyrimidines, purines, nucleosides, lipids and mixtures thereof. The indicator group is selected from compounds which have a first state when joined to the leaving group, and a second state when the leaving group is cleaved from the indicator group by the enzyme. Preferably, the indicator compounds are rhodamine 110, rhodol, and fluorescein and analogs of these compounds. A method of synthesizing the compound as well as methods of using these compounds to measure enzyme activity are also disclosed.

公开(公告)号：US06692702B1

公开(公告)日：2004-02-17

申请号：US09611847

申请日：2000-07-07

申请人： Alexander Burshteyn ,  John W. Joubran ,  Nazle Kuylen ,  Frank J. Lucas

发明人： Alexander Burshteyn ,  John W. Joubran ,  Nazle Kuylen ,  Frank J. Lucas

IPC分类号： G01N134

CPC分类号： G01N1/4077 ,  B01D61/147 ,  B01D61/18 ,  B01D61/20 ,  B01D63/02 ,  Y10T436/25375 ,  Y10T436/255

摘要： A method for utilizing a filtration device for removing interferants from a sample containing cells in an automated apparatus is disclosed. The filtration device includes a microporous hollow fiber membrane having a plurality of pores sized to retain cells while allowing smaller diameter interferants to pass through the membrane. The apparatus also includes a means for of moving the sample from a sample container to and from the filtration device. The disclosed method utilizes a vacuum source to aspirate the sample into a lumen of the hollow fiber membrane so that the sample is retained in the lumen space until expelled into an analysis container or transported to an analyzer.

摘要翻译： 公开了一种利用过滤装置从自动化装置中含有细胞的样品中除去干扰物的方法。 过滤装置包括具有多个孔的微孔中空纤维膜，其尺寸设计成保持细胞同时允许较小直径的干燥剂通过膜。 该设备还包括用于将样品从样品容器移动到过滤装置和从过滤装置移动的装置。 所公开的方法利用真空源将样品吸入中空纤维膜的内腔，使得样品保留在管腔空间中，直到排出到分析容器中或运送到分析器中。

公开(公告)号：US5776720A

公开(公告)日：1998-07-07

申请号：US443776

申请日：1995-05-18

申请人： Gerald E. Jaffe ,  Frank J. Lucas ,  James H. Carter

发明人： Gerald E. Jaffe ,  Frank J. Lucas ,  James H. Carter

IPC分类号： C12Q1/02 ,  C12Q1/34 ,  G01N33/574 ,  C12Q1/00

CPC分类号： C12Q1/34 ,  C12Q1/02 ,  G01N33/574 ,  C12Q2334/40 ,  C12Q2337/40

摘要： An assay compound or a salt thereof for assaying the activity of an enzyme inside a metabolically active whole cell is disclosed. The assay compound includes a leaving group and an indicator group. The leaving group is selected from the group comprising amino acids, peptides, saccharides, sulfates, phosphates, esters, phosphate esters, nucleotides, polynucleotides, nucleic acids, pyrimidines, purines, nucleosides, lipids and mixtures thereof. The indicator group is selected from compounds which have a first state when joined to the leaving group, and a second state when the leaving group is cleaved from the indicator group by the enzyme. Preferably, the indicator compounds are rhodamine 110, rhodol, and fluorescein and analogs of these compounds. A method of synthesizing the compound as well as methods of using these compounds to measure enzyme activity are also disclosed.

摘要翻译： 公开了一种用于测定代谢活性整个细胞内的酶的活性的测定化合物或其盐。 测定化合物包括离去基团和指示剂组。 离去基团选自包括氨基酸，肽，糖，硫酸盐，磷酸盐，酯，磷酸酯，核苷酸，多核苷酸，核酸，嘧啶，嘌呤，核苷，脂质及其混合物的组。 指示基团选自与离去基团连接时具有第一状态的化合物，当离去基团被酶从指示剂基团切割时的第二种状态。 优选地，指示剂化合物是这些化合物的罗丹明110，rhodol和荧光素以及类似物。 还公开了合成化合物的方法以及使用这些化合物测定酶活性的方法。

公开(公告)号：US5733719A

公开(公告)日：1998-03-31

申请号：US445217

申请日：1995-05-18

申请人： Gerald E. Jaffe ,  Frank J. Lucas ,  James H. Carter

发明人： Gerald E. Jaffe ,  Frank J. Lucas ,  James H. Carter

IPC分类号： C12Q1/34 ,  G01N33/574 ,  C12Q1/00 ,  C12Q1/02

CPC分类号： C12Q1/34 ,  G01N33/574 ,  Y10S435/968

摘要： An assay compound or a salt thereof for assaying the activity of an enzyme inside a metabolically active whole cell is disclosed. The assay compound includes a leaving group and an indicator group. The leaving group is selected from the group comprising amino acids, peptides, saccharides, sulfates, phosphates, esters, phosphate esters, nucleotides, polynucleotides, nucleic acids, pyrimidines, purines, nucleosides, lipids and mixtures thereof. The indicator group is selected from compounds which have a first state when joined to the leaving group, and a second state when the leaving group is cleaved from the indicator group by the enzyme. Preferably, the indicator compounds are rhodamine 110, rhodol, and fluorescein and analogs of these compounds. A method of synthesizing the compound as well as methods of using these compounds to measure enzyme activity are also disclosed.

摘要翻译： 公开了一种用于测定代谢活性整个细胞内的酶的活性的测定化合物或其盐。 测定化合物包括离去基团和指示剂组。 离去基团选自包括氨基酸，肽，糖，硫酸盐，磷酸盐，酯，磷酸酯，核苷酸，多核苷酸，核酸，嘧啶，嘌呤，核苷，脂质及其混合物的组。 指示基团选自与离去基团连接时具有第一状态的化合物，当离去基团被酶从指示剂基团切割时的第二种状态。 优选地，指示剂化合物是这些化合物的罗丹明110，rhodol和荧光素以及类似物。 还公开了合成化合物的方法以及使用这些化合物测定酶活性的方法。

公开(公告)号：US5871946A

公开(公告)日：1999-02-16

申请号：US444056

申请日：1995-05-18

申请人： Frank J. Lucas ,  Gerald E. Jaffe ,  Steven Bott ,  James H. Carter

发明人： Frank J. Lucas ,  Gerald E. Jaffe ,  Steven Bott ,  James H. Carter

IPC分类号： C12Q1/34 ,  G01N33/574 ,  C12Q1/02

CPC分类号： C12Q1/34 ,  G01N33/574 ,  Y10S435/962 ,  Y10S435/968

摘要： An assay compound or a salt thereof for assaying the activity of an enzyme inside a metabolically active whole cell is disclosed. The assay compound includes a leaving group and an indicator group. The leaving group is selected from the group comprising amino acids, peptides, saccharides, sulfates, phosphates, esters, phosphate esters, nucleotides, polynucleotides, nucleic acids, pyrimidines, purines, nucleosides, lipids and mixtures thereof. The indicator group is selected from compounds which have a first state when joined to the leaving group, and a second state when the leaving group is cleaved from the indicator group by the enzyme. Preferably, the indicator compounds are rhodamine 110, rhodol, and fluorescein and analogs of these compounds. A method of synthesizing the compound as well as methods of using these compounds to measure enzyme activity are also disclosed.

摘要翻译： 公开了一种用于测定代谢活性整个细胞内的酶的活性的测定化合物或其盐。 测定化合物包括离去基团和指示剂组。 离去基团选自包括氨基酸，肽，糖，硫酸盐，磷酸盐，酯，磷酸酯，核苷酸，多核苷酸，核酸，嘧啶，嘌呤，核苷，脂质及其混合物的组。 指示基团选自与离去基团连接时具有第一状态的化合物，当离去基团被酶从指示剂基团切割时的第二种状态。 优选地，指示剂化合物是这些化合物的罗丹明110，rhodol和荧光素以及类似物。 还公开了合成化合物的方法以及使用这些化合物测定酶活性的方法。