公开(公告)号：US5494793A

公开(公告)日：1996-02-27

申请号：US366971

申请日：1989-06-14

申请人： Deborah C. Schindele ,  Barry V. Pepich ,  George E. Renzoni ,  Karen L. Fearon ,  Niels H. Andersen ,  Thomas H. Stanton

发明人： Deborah C. Schindele ,  Barry V. Pepich ,  George E. Renzoni ,  Karen L. Fearon ,  Niels H. Andersen ,  Thomas H. Stanton

IPC分类号： A61K41/00 ,  C07H21/00 ,  C12Q1/68 ,  G01N33/533 ,  C07H21/04 ,  C12N9/00 ,  G01N33/53

CPC分类号： G01N33/533 ,  A61K41/0071 ,  C07H21/00 ,  C12Q1/6816 ,  C12Q1/6869

摘要： Fluorescent and/or chromogenic reagents in which a phthalocyanine derivative is monomerically conjugated with an antigen, antibody, oligonucleotide, or nucleic acid. Methods are presented in in which greater than 90% of the phthalocyanine dyes are monomeric when conjugated. This greatly enhances their performance as detectable markers in immunoassays, nucleic acid probe assays, immunoblotting, hybridization assays, microscopy, imaging, flow cytometry, DNA sequencing, and photodynamic therapy. For use as fluorophores, the free base phthalocyanine may or may not be metallated. Metals for fluorescent phthalocyanine include aluminum, silicon, phosphorus, gallium, germanium, cadmium, scandium, magnesium, tin, and zinc. For use as chromogens, the phthalocyanine may or may not be metallated. For use in aqueous solution, the phthalocyanine macrocycle should be derivatized with water-solubilizing substituents such as sulfonic acid, phosphate, phosphonate, hydroxy, phenoxy, amino, ammonium, or pyridinium groups. To promote disaggregation, metallation with an atom of +3 valence or higher is recommended, so that the monomer will take on an axial ligand in aqueous solution. For use in enzymatic immunoassays and enzymatically enhanced nucleic acid probe assays, the monomeric phthalocyanine derivative is conjugated via an enzyme-cleavable linkage with the antigen, antibody, oligonucleotide, or nucleic acid. Reversibly quenched embodiments are also provided in which a cleavable linkage joins a fluorescent phthalocyanine monomer with another phthalocyanine, a heavy metal, or a paramagnetic species.

摘要翻译： 荧光和/或显色试剂，其中酞菁衍生物与抗原，抗体，寡核苷酸或核酸单体缀合。 其中存在大于90％的酞菁染料当共轭时是单体的方法。 这大大提高了其作为免疫测定中的可检测标记，核酸探针测定，免疫印迹，杂交测定，显微镜，成像，流式细胞术，DNA测序和光动力学治疗的性能。 为了用作荧光团，游离碱酞菁可以是或可以不是金属化的。 荧光酞菁的金属包括铝，硅，磷，镓，锗，镉，钪，镁，锡和锌。 为了用作色素，酞菁可以是或可以不是金属化的。 为了在水溶液中使用，酞菁大环化合物应当用水溶性取代基如磺酸，磷酸酯，膦酸酯，羟基，苯氧基，氨基，铵或吡啶鎓基衍生化。 为了促进分解，推荐使用+3价以上的原子进行金属化，使单体在水溶液中呈轴向配体。 为了用于酶免疫测定和酶促增强的核酸探针测定，单体酞菁衍生物通过酶切断连接与抗原，抗体，寡核苷酸或核酸缀合。 还提供了可逆淬火的实施方案，其中可切割键连接荧光酞菁单体与另一种酞菁，重金属或顺磁性物质。

公开(公告)号：US5135717A

公开(公告)日：1992-08-04

申请号：US398433

申请日：1989-08-29

申请人： George E. Renzoni ,  Deborah C. Schindele ,  Louis J. Theodore ,  Clifford C. Leznoff ,  Karen L. Fearon ,  Barry V. Pepich

发明人： George E. Renzoni ,  Deborah C. Schindele ,  Louis J. Theodore ,  Clifford C. Leznoff ,  Karen L. Fearon ,  Barry V. Pepich

IPC分类号： A61K41/00 ,  C07H21/00 ,  C12Q1/68 ,  G01N33/533 ,  G01N33/58

CPC分类号： C07H21/00 ,  A61K41/0071 ,  G01N33/533 ,  C12Q1/6869 ,  Y10S435/81 ,  Y10S435/975 ,  Y10S436/808 ,  Y10T436/143333

摘要： Red-shifted, water-soluble, fluorescent, monomerically-tetherable derivatives having the formula: ##STR1## wherein, M represents either H.sub.2 or is selected from among the following metals: aluminum, silicon, phosphorus, gallium, germanium, cadmium, scandium, magnesium, tin, and zinc. Each R.sub.1 is independently selected from --XYW, --YW, and --W. X represents either a carbon, or heteroatom selected from among oxygen, nitrogen, sulfur, phosphorus, silicon, and selenium; Y represents a linking group; and W represents a water solubilizing group. The substituent R.sub.2 is selected from among --A, --Y'A, --XA, and --XY'A, where A denotes a biological entity such as an antibody, antibody fragment, nucleotide, nucleic acid probe, antigen, oligonucleotide, deoxynucleotide, dideoxynucleotide, avidin, streptavidin or membrane probe, or R.sub.2 is a reactive or activatable group suitable for conjugating to a biological entity. Y' is a linking group that tethers the biological entity to the phthalocyanine or tetrabenztriazaporphyrin macrocycle. Z is either a nitrogen atom or a carbon substituted with hydrogen, alkyl, aryl, or aralkyl groups. Z may also be attached to R.sub.2. Also disclosed are derivatives of the compounds of the above formula in which 1-4 of the benzo rings(s) contain 1 or 2 N atoms. Methods of sequencing DNA and detecting analytes, including cells, using these derivatives are disclosed, as are kits for carrying out assays for the analytes and flow cytometry. Methods of detecting DNA using cationic compounds of the above formula, wherein R.sub.2 =R.sub.1 and W=--N.sup.+ D.sub.1 D.sub.2 D.sub.3 are also disclosed. Further, compounds containing Tc, Gd, etc. as the metal in the above formula may be used for imaging or therapy.

摘要翻译： 具有下式的红移，水溶性，荧光，单体 - 可连接的衍生物：其中，M表示H 2或选自下列金属：铝，硅，磷，镓，锗，镉，钪， 镁，锡和锌。 每个R1独立地选自-XYW，-YW和-W。 X表示选自氧，氮，硫，磷，硅和硒中的碳或杂原子; Y表示连接基团; W表示水溶性基团。 取代基R2选自-A，-Y'A，-XA和-XY'A，其中A表示生物实体，例如抗体，抗体片段，核苷酸，核酸探针，抗原，寡核苷酸，脱氧核苷酸， 双脱氧核苷酸，抗生物素蛋白，链霉抗生物素蛋白或膜探针，或R2是适于与生物实体缀合的反应性或活化性基团。 Y'是将生物体系到酞菁或四苯并三氮杂卟啉大环的连接基团。 Z是氮原子或被氢，烷基，芳基或芳烷基取代的碳。 Z也可以连接到R2。 还公开了上式化合物的衍生物，其中1-4个苯并环含有1或2个N原子。 公开了使用这些衍生物测序DNA和检测分析物（包括细胞）的方法，以及用于进行分析物和流式细胞术的测定的试剂盒。 还公开了使用上式的阳离子化合物检测DNA的方法，其中R2 = R1和W = -N + D1D2D3。 此外，可以使用含有上述式中的金属的Tc，Gd等的化合物进行成像或治疗。

公开(公告)号：US5346670A

公开(公告)日：1994-09-13

申请号：US895601

申请日：1992-06-08

申请人： George E. Renzoni ,  Deborah C. Schindele ,  Louis J. Theodore ,  Clifford C. Leznoff ,  Karen L. Fearon ,  Barry V. Pepich

发明人： George E. Renzoni ,  Deborah C. Schindele ,  Louis J. Theodore ,  Clifford C. Leznoff ,  Karen L. Fearon ,  Barry V. Pepich

IPC分类号： A61K41/00 ,  C07H21/00 ,  C12Q1/68 ,  G01N33/533 ,  G01N21/76

CPC分类号： G01N33/533 ,  A61K41/0071 ,  C07H21/00 ,  Y10S435/808 ,  Y10S435/968 ,  Y10S436/808 ,  Y10T436/13

摘要： Red-shifted, water-soluble, fluorescent, monomerically-tetherable derivatives having the formula: ##STR1##wherein, M represents either H.sub.2 or is selected from among the following metals: aluminum, silicon, phosphorus, gallium, germanium, cadmium, scandium, magnesium, tin, and zinc. Each R.sub.1 is independently selected from --XYW, --YW, and --W. X represents either a carbon, or heteroatom selected from among oxygen, nitrogen, sulfur, phosphorus, silicon, and selenium; Y represents a linking group; and W represents a water solubilizing group. The substituent R.sub.2 is selected from among --A, --Y'A, --XA, and --XY'A, where A denotes a biological entity such as an antibody, antibody fragment, nucleotide, nucleic acid probe, antigen, oligonucleotide, deoxynucleotide, dideoxynucleotide, avidin, streptavidin or membrane probe, or R.sub.2 is a reactive or activatable group suitable for conjugating to a biological entity. Y' is a linking group that tethers the biological entity to the phthalocyanine or tetrabenztriazaporphyrin macrocycle. Z is either a nitrogen atom or a carbon substituted with hydrogen, alkyl, aryl, or aralkyl groups. Z may also be attached to R.sub.2. Also disclosed are derivatives of the compounds of the above Formula in which 1-4 of the benzo ring(s) contain 1 or 2 N atoms. Methods of sequencing DNA and detecting analytes, including cells, using these derivatives are disclosed, as are kits for carrying out assays for the analytes and flow cytometry. Methods of detecting DNA using cationic compounds of the above Formula, wherein R.sub.2 =R.sub.1 and W=-N.sup.+ D.sub.1 D.sub.2 D.sub.3 are also disclosed. Further, compounds containing Tc, Gd, etc. as the metal in the above Formula may be used for imaging or therapy.

摘要翻译： 具有下式的红移，水溶性，荧光，单体 - 可连接的衍生物：选自以下金属：铝，硅，磷，镓，锗，镉，钪，镁，锡和锌。 每个R1独立地选自-XYW，-YW和-W。 X表示选自氧，氮，硫，磷，硅和硒中的碳或杂原子; Y表示连接基团; W表示水溶性基团。 取代基R2选自-A，-Y'A，-XA和-XY'A，其中A表示生物实体，例如抗体，抗体片段，核苷酸，核酸探针，抗原，寡核苷酸，脱氧核苷酸， 双脱氧核苷酸，抗生物素蛋白，链霉抗生物素蛋白或膜探针，或R2是适于与生物实体缀合的反应性或活化性基团。 Y'是将生物实体连接到酞菁或四苯并三氮杂卟啉大环的连接基团。 Z是氮原子或被氢，烷基，芳基或芳烷基取代的碳。 Z也可以连接到R2。 还公开了上述式的化合物的衍生物，其中1-4个苯并环含有1或2个N原子。 公开了使用这些衍生物测序DNA和检测分析物（包括细胞）的方法，以及用于进行分析物和流式细胞术的测定的试剂盒。 还公开了使用上述式的阳离子化合物（其中R2 = R1和W = -N + D1D2D3）检测DNA的方法。 此外，可以使用含有上述式中的金属的Tc，Gd等的化合物进行成像或治疗。

公开(公告)号：US4803170A

公开(公告)日：1989-02-07

申请号：US946475

申请日：1986-12-24

申请人： Thomas H. Stanton ,  James A. Clagett ,  Deborah C. Schindele ,  George E. Renzoni ,  Niels H. Andersen ,  Kent E. Opheim

发明人： Thomas H. Stanton ,  James A. Clagett ,  Deborah C. Schindele ,  George E. Renzoni ,  Niels H. Andersen ,  Kent E. Opheim

IPC分类号： G01N33/533 ,  G01N33/543 ,  G01N33/558 ,  G01N33/53

CPC分类号： G01N33/54306 ,  G01N33/533 ,  G01N33/558 ,  Y10S436/807 ,  Y10S436/808 ,  Y10S436/809

摘要： An immunoassay device including one or more reaction chambers, each adapted to receive and retain a volume of test fluid in fluid communication with nonoverlapping first, second, and third reagent-bearing surfaces. To the first surface is reversibly bound an analyte conjugate: analyte component conjugated to one or more components, termed ligand/marker, that serve ligand and marker functions as described herein. Analyte binding partner is immobilized on the second surface, and ligand/marker binding partner is immobilized on the third surface. The reaction chamber is preferably configured to receive and direct the test fluid sequentially past the first second, and third reagent surfaces. In use, analyte conjugate solubilized from the first surface completes with any analyte in the test fluid for analyte binding partner sites on the second surface. Excess analyte conjugate becomes sequestered on the third surface, where the marker activity is read to indicate analyte presence and concentration in the test fluid. A test kit includes the immunoassay device in combination with comparative test results.

摘要翻译： 一种包括一个或多个反应室的免疫测定装置，每个反应室适于容纳和保持与非重叠的第一，第二和第三试剂载体表面流体连通的一定体积的测试流体。 第一表面可逆地结合分析物缀合物：与一种或多种组分缀合的分析物成分，称为配体/标记，其用于如本文所述的配体和标记功能。 分析物结合配偶体固定在第二表面上，并且配体/标记结合配偶体固定在第三表面上。 反应室优选地构造成接收并引导测试流体依次经过第一和第三试剂表面。 在使用中，从第一表面溶解的分析物结合物与第二表面上分析物结合配偶位点的测试流体中的任何分析物完成。 超量分析物缀合物在第三表面上被隔离，其中读取标记活性以指示分析物在测试流体中的存在和浓度。 测试试剂盒包括免疫测定装置和比较试验结果。