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1. 发明申请

US20060148023A1 Affinity labeling of serine proteases for simultaneous detection of multiple serine protease activity levels 审中-公开
标题翻译：亲和标记丝氨酸蛋白酶同时检测多个丝氨酸蛋白酶活性水平
公开(公告)号：US20060148023A1
公开(公告)日：2006-07-06
申请号：US10872824
申请日：2004-06-21
申请人： David Phelps , Gary Johnson , Brian Lee , Zbigniew Darzynkiewicz , Jerzy Grabarek
发明人： David Phelps , Gary Johnson , Brian Lee , Zbigniew Darzynkiewicz , Jerzy Grabarek
IPC分类号： C12Q1/37
CPC分类号： C12Q1/37 , G01N33/5091 , G01N33/573 , G01N2500/04 , G01N2510/00 , G01N2800/52
摘要： The invention provides assay methods and reagents useful for evaluating the level of enzyme activities within living cells. Enzyme activity levels within living cells, such as Serine proteases, can be key determinates in assessing; 1) the presence of tumor (cancer) cells, 2) the predictive efficacy of a chemotherapeutic treatment regimen using a particular therapeutic agent or process, 4) the probability of graft rejection or acceptance, and 5) the disease state status of a cell. The identification of the up or down regulation relationships of serine proteases within living cell systems, provides a rapid, yet finely tuned mechanism for predicting the current and future physiological state of these cell populations.
摘要翻译：本发明提供了可用于评估活细胞内酶活性水平的测定方法和试剂。活细胞内的酶活性水平，如丝氨酸蛋白酶，可能是评估中的关键决定因素; 1）肿瘤（癌症）细胞的存在，2）使用特定治疗剂或过程的化学治疗方案的预测效果，4）移植排斥或接受的概率，以及5）细胞的疾病状态。识别活细胞系统中丝氨酸蛋白酶的上调或下调调节关系，为预测这些细胞群体的当前和未来生理状态提供了一种快速但精细调节的机制。

2. 发明申请

US20050136492A1 Affinity labeling of enzymes for detection of enzyme activity level in living cells 审中-公开
标题翻译：用于检测活细胞中酶活性水平的酶的亲和标记
公开(公告)号：US20050136492A1
公开(公告)日：2005-06-23
申请号：US10872818
申请日：2004-06-21
申请人： David Phelps , Gary Johnson , Brian Lee , Zbigniew Darzynkiewicz , Jerzy Grabarek
发明人： David Phelps , Gary Johnson , Brian Lee , Zbigniew Darzynkiewicz , Jerzy Grabarek
IPC分类号： C12Q1/37 , G01N33/573 , G01N33/574
CPC分类号： G01N33/573 , C12Q1/37 , G01N33/574
摘要： The invention provides assay methods and reagents useful for evaluating the level of enzyme activities within living cells. Enzyme activity levels within living cells, such as caspases and Serine proteases, can be key determinates in assessing; 1) the apoptotic state of a cell, 2) the presence of tumor (cancer) cells, 3) the predictive efficacy of a chemotherapeutic treatment regimen using a particular therapeutic agent or process, 4) the probability of graft rejection or acceptance, identification of the up or down regulation relationships of serine proteases and caspases within living cell systems, provides a rapid, yet finely tuned mechanism for predicting the current and future state of these cell populations, and 5) the disease state status of a cell.
摘要翻译：本发明提供了可用于评估活细胞内酶活性水平的测定方法和试剂。活细胞内的酶活性，如胱天蛋白酶和丝氨酸蛋白酶，可以作为评估中的关键决定因素。 1）细胞的凋亡状态，2）肿瘤（癌症）细胞的存在，3）使用特定治疗剂或过程的化学治疗方案的预测效果，4）移植排斥或接受的概率，鉴定丝氨酸蛋白酶和半胱氨酸蛋白酶在活细胞系统中的向上或向下调节关系为预测这些细胞群体的当前和未来状态提供了一种快速但精细调节的机制，以及5）细胞的疾病状态。

3. 发明申请

US20070269832A1 Novel affinity labels 审中-公开
标题翻译：新颖的亲和标签
公开(公告)号：US20070269832A1
公开(公告)日：2007-11-22
申请号：US10872754
申请日：2004-06-21
申请人： David Phelps , Gary Johnson , Brian Lee , Zbigniew Darzynkiewicz , Jerzy Grabarek
发明人： David Phelps , Gary Johnson , Brian Lee , Zbigniew Darzynkiewicz , Jerzy Grabarek
IPC分类号： C12Q1/02 , C07D311/82 , C12Q1/37 , G01N33/573
CPC分类号： C07K5/06191 , C12Q1/37 , G01N33/58 , G01N2500/04
摘要： The invention provides novel affinity labels for Ser proteases of formula: L-A-X—NH—CH(R′)C(═O)CH2Cl (I) wherein L, A, X, and R′ have any of the values defined in the specification, or salts thereof, as well as compositions comprising such compounds or salts. The composition of the amino acid side-chain (R′) along with the amino acid or amino acid sequence (peptide) of the X component of formula I, affect the target selectivity of the labeled affinity ligand. Utilization of cell permeable, enzyme selective, labeled affinity ligands, provides a precise mechanism for evaluating the current and future status of cell populations.
摘要翻译：本发明提供了用于下式的Ser蛋白酶的新型亲和标记：<？in-line-formula description =“In-line Formulas”end =“lead”？> LAX-NH-CH（R'）C（-O）CH 其中L，A，X和R'具有如下定义的任何值：（1）说明书或其盐，以及包含这些化合物或盐的组合物。氨基酸侧链（R'）的组成与式I的X组分的氨基酸或氨基酸序列（肽）一起影响标记的亲和配体的靶选择性。细胞可渗透，酶选择性，标记的亲和配体的利用提供了一种精确的机制来评估细胞群体的当前和未来状态。

4. 发明授权

US06417343B1 Physical separation of nucleic acids by antibodies to halogenated nucleotides 失效
标题翻译：通过抗体与卤代核苷酸物理分离核酸
公开(公告)号：US06417343B1
公开(公告)日：2002-07-09
申请号：US08951598
申请日：1997-10-16
申请人： Zbigniew Darzynkiewicz , Frank Traganos , Gloria Juan
发明人： Zbigniew Darzynkiewicz , Frank Traganos , Gloria Juan
IPC分类号： C12P1934
CPC分类号： C12Q1/6804 , G01N33/5308 , C12Q2525/117
摘要： Methods for immunocytochemical detection and isolation of RNA.
摘要翻译：免疫细胞化学检测和RNA分离方法

5. 发明申请

US20050042694A1 Detection of protein conformations in single cells 失效
标题翻译：单细胞中蛋白质构象的检测
公开(公告)号：US20050042694A1
公开(公告)日：2005-02-24
申请号：US10954097
申请日：2004-09-29
申请人： Zbigniew Darzynkiewicz , Frank Traganos , Gloria Juan , Stefan Gruenwald
发明人： Zbigniew Darzynkiewicz , Frank Traganos , Gloria Juan , Stefan Gruenwald
IPC分类号： G01N33/50 , G01N33/53 , G01N33/543 , G01N33/574 , G01N33/68 , C07K16/46 , G01N33/567
CPC分类号： G01N33/6845 , G01N33/5011 , G01N33/53 , G01N33/543 , G01N33/574 , G01N33/68 , G01N33/6842 , G01N2333/4736 , G01N2333/9121 , Y10S435/973 , Y10S435/974 , Y10S436/80 , Y10S436/813
摘要： Methods, reagents, and kits are provided that permit flow cytometric determination of the phosphorylation status of retinoblastoma susceptibility gene protein (pRB) in individual cells. Methods are described that permit the hypophosphorylated, active, form of pRB to be measured either as an absolute quantity or as a proportion of total cellular pRB. Further described are methods that permit pRB phosphorylation status to be correlated with cell cycle phase and with protein components of the cell cycle. Screening of chemical compounds for antiproliferative and antineoplastic activity using the flow cytometric assays is demonstrated. Reagent kits that facilitate the subject methods are also provided.
摘要翻译：提供方法，试剂和试剂盒，其允许流式细胞术测定个体细胞中视网膜母细胞瘤易感性基因蛋白（pRB）的磷酸化状态。描述允许磷酸化，活性形式的pRB作为绝对量或总细胞pRB的比例来测量的方法。进一步描述的是允许pRB磷酸化状态与细胞周期相关和细胞周期的蛋白质组分相关联的方法。证明使用流式细胞术检测筛选抗增殖和抗肿瘤活性的化合物。还提供了促进主题方法的试剂盒。

6. 发明授权

US4559309A Flow cytometry-fluorescence measurements for characterizing sperm 失效
标题翻译：用于表征精子的流式细胞术 - 荧光测量
公开(公告)号：US4559309A
公开(公告)日：1985-12-17
申请号：US413862
申请日：1982-09-01
申请人： Donald P. Evenson , Zbigniew Darzynkiewicz
发明人： Donald P. Evenson , Zbigniew Darzynkiewicz
IPC分类号： G01N1/30 , G01N15/14 , G01N21/64 , G01N33/52
CPC分类号： G01N21/6428 , G01N1/30 , G01N15/14 , G01N2021/6421 , Y10T436/143333
摘要： A process for characterizing sperm motility and viability by staining a sperm sample with Rhodamine 123 and ethidium bromide, and simultaneously measuring the sperm fluorescence emissions at green frequencies 515-575 nm and at red frequencies 600-650 nm, the green counts being correlated with sperm motility and the red counts being correlated with putative dying or dead cells. Additionally, a sample of sperm can be characterized as to type and normality by staining a sample of sperm with acridine orange and simultaneously measuring the sperm fluorescence emissions at green frequencies 515-575 nm and at red frequencies 600-650 nm.
摘要翻译：通过用罗丹明123和溴化乙锭染色精子样品来表征精子活力和活力的过程，同时测量在515-575nm的绿色频率和在600-650nm的红色频率下的精子荧光发射，绿色计数与精子相关运动性和红色计数与假定的死亡或死亡细胞相关。此外，精子样品可以通过用吖啶橙染色精子样品并同时测量在515-575nm的绿色频率和在600-650nm的红色频率下的精子荧光发射，来表征其类型和正常性。

7. 发明申请

US20110214680A1 APPROACHES TO IDENTIFY LESS HARMFUL TOBACCO AND TOBACCO PRODUCTS 审中-公开
标题翻译：鉴定没有有害烟草和烟草制品的方法
公开(公告)号：US20110214680A1
公开(公告)日：2011-09-08
申请号：US13073811
申请日：2011-03-28
申请人： Anthony P. Albino , Ellen D. Jorgensen , Frank Traganos , Zbigniew Darzynkiewicz , Wendy Jin
发明人： Anthony P. Albino , Ellen D. Jorgensen , Frank Traganos , Zbigniew Darzynkiewicz , Wendy Jin
IPC分类号： A24C5/32 , C12Q1/68
CPC分类号： A01H1/04 , C12N9/0004 , C12N15/8227 , C12N15/8243 , C12N15/8274
摘要： Aspects of the invention concern methods for detecting, identifying and evaluating tobacco and tobacco products to determine the potential that these compositions have to contribute to a tobacco-related disease. It is based, at least in part; on the discovery that exposure of pulmonary cells to smoke or smoke condensate obtained from tobacco or tobacco products induces double stranded breaks in cellular DNA, which were efficiently detected using assays that measure the presence, absence, or amount of phosphorylation of the histone, H2AX.
摘要翻译：本发明的方面涉及用于检测，鉴定和评估烟草和烟草制品以确定这些组合物对烟草相关疾病有潜力的方法。它基于，至少部分; 发现肺细胞暴露于从烟草或烟草制品获得的烟雾或烟雾的冷凝物诱导细胞DNA中的双链断裂，其使用测量组蛋白H2AX的磷酸化的存在，不存在或量的测定法被有效检测。

8. 发明授权

US07662565B2 Approaches to identify less harmful tobacco and tobacco products 有权
标题翻译：识别较少有害烟草和烟草制品的方法
公开(公告)号：US07662565B2
公开(公告)日：2010-02-16
申请号：US11596088
申请日：2005-05-11
申请人： Anthony P. Albino , Ellen D. Jorgensen , Frank Traganos , Zbigniew Darzynkiewicz , Wendy Jin
发明人： Anthony P. Albino , Ellen D. Jorgensen , Frank Traganos , Zbigniew Darzynkiewicz , Wendy Jin
IPC分类号： C12Q1/68 , C12Q1/02
CPC分类号： A01H1/04 , C12N9/0004 , C12N15/8227 , C12N15/8243 , C12N15/8274
摘要： Aspects of the invention concern methods for detecting, identifying and evaluating tobacco and tobacco products to determine the potential that these compositions have to contribute to a tobacco-related disease. It is based, at least in part; on the discovery that exposure of pulmonary cells to smoke or smoke condensate obtained from tobacco or tobacco products induces double stranded breaks in cellular DNA, which were efficiently detected using assays that measure the presence, absence, or amount of phosphorylation of the histone, H2AX.
摘要翻译：本发明的方面涉及用于检测，鉴定和评估烟草和烟草制品以确定这些组合物对烟草相关疾病有潜力的方法。它基于，至少部分; 发现肺细胞暴露于从烟草或烟草制品获得的烟雾或烟雾的冷凝物诱导细胞DNA中的双链断裂，其使用测量组蛋白H2AX的磷酸化的存在，不存在或量的测定法被有效检测。

9. 发明授权

US06821740B2 Flow cytometric methods for the concurrent detection of discrete functional conformations of PRB in single cells 有权
标题翻译：用于在单细胞中同时检测PRB的离散功能构象的流式细胞术方法
公开(公告)号：US06821740B2
公开(公告)日：2004-11-23
申请号：US09256817
申请日：1999-02-24
申请人： Zbigniew Darzynkiewicz , Frank Traganos , Gloria Juan , Stefan Gruenwald
发明人： Zbigniew Darzynkiewicz , Frank Traganos , Gloria Juan , Stefan Gruenwald
IPC分类号： G01N33574
CPC分类号： G01N33/6845 , G01N33/5011 , G01N33/53 , G01N33/543 , G01N33/574 , G01N33/68 , G01N33/6842 , G01N2333/4736 , G01N2333/9121 , Y10S435/973 , Y10S435/974 , Y10S436/80 , Y10S436/813
摘要： Methods, reagents, and kits are provided that permit flow cytometric determination of the phosphorylation status of retinoblastoma susceptibility gene protein (pRB) in individual cells. Methods are described that permit the hypophosphorylated, active, form of pRB to be measured either as an absolute quantity or as a proportion of total cellular pRB. Further described are methods that permit pRB phosphorylation status to be correlated with cell cycle phase and with protein components of the cell cycle. Screening of chemical compounds for antiproliferative and antineoplastic activity using the flow cytometric assays is demonstrated. Reagent kits that facilitate the subject methods are also provided.
摘要翻译：提供方法，试剂和试剂盒，其允许流式细胞术测定个体细胞中视网膜母细胞瘤易感性基因蛋白（pRB）的磷酸化状态。描述允许磷酸化，活性形式的pRB作为绝对量或总细胞pRB的比例来测量的方法。进一步描述的是允许pRB磷酸化状态与细胞周期相关和细胞周期的蛋白质组分相关联的方法。证明使用流式细胞术检测筛选抗增殖和抗肿瘤活性的化合物。还提供了促进主题方法的试剂盒。

10. 发明授权

US5912126A Methods for labeling DNA ends with halogenated nucleotides and detecting same with antibodies 失效
标题翻译：用卤代核苷酸标记DNA末端并用抗体检测DNA的方法
公开(公告)号：US5912126A
公开(公告)日：1999-06-15
申请号：US727509
申请日：1996-10-22
申请人： Zbigniew Darzynkiewicz , Xun Li , Frank Traganos
发明人： Zbigniew Darzynkiewicz , Xun Li , Frank Traganos
IPC分类号： C07H21/00 , C07K16/44 , C12Q1/68
CPC分类号： C12Q1/6804 , C07H21/00 , C07K16/44 , C12Q1/6816 , C12Q1/6841
摘要： The invention pertains to the field of DNA detection for basic research, medical diagnostic testing, and forensic testing. Methods are provided for end labeling of DNA strands without a denaturation step so that cellular morphology can be better preserved. The DNA strands are first incubated with a halogenated deoxynucleotide triphosphate, such as brominated deoxyuridine triphosphate (BrdUTP), and an enzyme which can catalyze the addition of the halogenated deoxynucleotide to the 3' OH ends of the DNA strand, such as terminal deoxynucleotidyl transferase (TdT). The resulting modified DNA strands are then incubated with a labeled antibody, such as a fluoresceinated monoclonal antibody, that binds specifically to the halogenated deoxynucleotide. The label is then detected, e.g., by flow cytometry. The methods have utility in detecting apotosis, DNA synthesis and/or repair, and as general methods for end labeling DNA.
摘要翻译： PCT No.PCT / US96 / 14250 Sec。 371日期1996年10月22日第 102（e）日期1996年10月22日PCT提交1996年8月28日PCT公布。公开号WO97 / 08345 日期1997年3月6日本发明涉及基础研究，医学诊断测试和法医检测的DNA检测领域。提供了用于DNA链末端标记而不进行变性步骤的方法，使得可以更好地保留细胞形态。首先将DNA链与卤化脱氧核苷酸三磷酸（例如溴化脱氧尿苷三磷酸（BrdUTP））一起温育，以及可以催化向DNA链的3'OH末端添加卤代脱氧核苷酸的酶，例如末端脱氧核苷酸转移酶 TdT）。然后将得到的修饰的DNA链与特异性结合卤代脱氧核苷酸的标记抗体如荧光单克隆抗体孵育。然后例如通过流式细胞术检测标记。该方法可用于检测凋亡，DNA合成和/或修复，以及作为终末标记DNA的一般方法。

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