公开(公告)号：WO1995015981A1

公开(公告)日：1995-06-15

申请号：PCT/US1994006166

申请日：1994-06-01

Applicant: UNIVERSITY OF UTAH RESEARCH FOUNDATION

Inventor： UNIVERSITY OF UTAH RESEARCH FOUNDATION ,  WEI, Ai-Ping ,  HERRON, James, W.

IPC: C07K15/12

CPC classification number: C07K14/59 ,  G01N33/542

Abstract: An improved oligopeptide composition for use in a fluorescent polarization immunoassay for a high molecular weight analyte is disclosed, along with a kit and a method using the composition. The composition comprises an oligopeptide selected by a screening procedure in which a plurality of different oligopeptides having respective amino acid sequences that represent sequential overlapping segments of the analyte amino acid sequence, and a fluorescent label bound thereto. The screening process further includes screening to select a composition which exhibits fluorescence enhancement upon binding the analyte. A preferred embodiment of the oligopeptide is one having an amino acid sequence which does not form internal disulfide bridges. Such a preferred oligopeptide will generally have no more than one cysteine residue. In a further preferred embodiment, the fluorescent label is tetramethylrhodamine or a cyanine dye. The kit may be packaged with instructions directing a user to prepare an assay solution containing the monoclonal antibody and the oligopeptide in certain respective concentrations. The composition, method and kit are constructed to detect nanomolar concentrations of the analyte.

Abstract translation: 公开了用于高分子量分析物的荧光偏振免疫测定法的改进的寡肽组合物以及试剂盒和使用该组合物的方法。 该组合物包含通过筛选方法选择的寡肽，其中多个具有代表分析物氨基酸序列的顺序重叠片段的氨基酸序列的不同寡肽和与其结合的荧光标记。 筛选方法还包括筛选以选择在结合分析物时显示荧光增强的组合物。 寡肽的优选实施方案是具有不形成内部二硫键的氨基酸序列的寡肽。 这种优选的寡肽通常具有不超过一个半胱氨酸残基。 在另一优选实施方案中，荧光标记是四甲基罗丹明或花青染料。 试剂盒可以包装，指示用户在某些相应浓度下制备含有单克隆抗体和寡肽的测定溶液。 构建组合物，方法和试剂盒以检测分析物的纳摩尔浓度。

公开(公告)号：WO1995003429A1

公开(公告)日：1995-02-02

申请号：PCT/US1994008279

申请日：1994-07-22

Applicant: UNIVERSITY OF UTAH RESEARCH FOUNDATION

Inventor： UNIVERSITY OF UTAH RESEARCH FOUNDATION ,  WEI, Ai-Ping ,  HERRON, James, N.

IPC: C12Q01/68

CPC classification number: G01N33/582 ,  C12Q1/6818 ,  Y10S436/80 ,  Y10S436/805

Abstract: Fluorescent energy transfer dyes capable of moving between a more stacked configuration to exhibit fluorescent quenching and a more spaced configuration to exhibit fluorescence can be conjugated to a peptide epitope or nucleic acid for use in the detection of an unknown antibody in bulk solution. The resulting labelled peptide reagent can be used in an immunoassay procedure by placing it in bulk solution along with the unknown antibody to be detected. When the antibody binds to the peptide epitope, the pair of dyes carried by the peptide epitope will have their configuration altered from a stacked to an unstacked configuration and will exhibit a fluorescent increase in response thereto.

Abstract translation: 能够在更叠层的配置之间移动以展现荧光猝灭和更间隔的配置以显示荧光的荧光能量转移染料可以缀合到肽表位或核酸上，以用于检测本体溶液中的未知抗体。 所得标记的肽试剂可以在免疫测定程序中使用，将其与待检测的未知抗体一起放入本体溶液中。 当抗体与肽表位结合时，由肽表位携带的一对染料将具有从堆叠到未堆叠构型的改变，并且对其反应将呈现荧光增加。

公开(公告)号：EP0733063B1

公开(公告)日：2003-12-17

申请号：EP94922433.1

申请日：1994-06-01

Applicant: UNIVERSITY OF UTAH RESEARCH FOUNDATION

Inventor： WEI, Ai-Ping ,  HERRON, James W.

IPC: C07K14/435 ,  C07K16/00 ,  G01N33/53 ,  G01N33/533 ,  G01N33/542 ,  G01N33/543 ,  G01N33/76 ,  G01N33/58

CPC classification number: C07K14/59 ,  G01N33/542

公开(公告)号：EP0733063A1

公开(公告)日：1996-09-25

申请号：EP94922433.0

申请日：1994-06-01

Applicant: UNIVERSITY OF UTAH RESEARCH FOUNDATION

Inventor： WEI, Ai-Ping ,  HERRON, James, W.

IPC: C07K14 ,  G01N33

CPC classification number: C07K14/59 ,  G01N33/542

Abstract: An improved oligopeptide composition for use in a fluorescent polarization immunoassay for a high molecular weight analyte is disclosed, along with a kit and a method using the composition. The composition comprises an oligopeptide selected by a screening procedure in which a plurality of different oligopeptides having respective amino acid sequences that represent sequential overlapping segments of the analyte amino acid sequence, and a fluorescent label bound thereto. The screening process further includes screening to select a composition which exhibits fluorescence enhancement upon binding the analyte. A preferred embodiment of the oligopeptide is one having an amino acid sequence which does not form internal disulfide bridges. Such a preferred oligopeptide will generally have no more than one cysteine residue. In a further preferred embodiment, the fluorescent label is tetramethylrhodamine or a cyanine dye. The kit may be packaged with instructions directing a user to prepare an assay solution containing the monoclonal antibody and the oligopeptide in certain respective concentrations. The composition, method and kit are constructed to detect nanomolar concentrations of the analyte.

公开(公告)号：EP0713534B1

公开(公告)日：2002-05-02

申请号：EP94922687.2

申请日：1994-07-22

Applicant: University of Utah Research Foundation

Inventor： WEI, Ai-Ping ,  HERRON, James N.

IPC: C12Q1/68 ,  G01N33/542

CPC classification number: G01N33/582 ,  C12Q1/6818 ,  Y10S436/80 ,  Y10S436/805

公开(公告)号：EP0713534A1

公开(公告)日：1996-05-29

申请号：EP94922687.0

申请日：1994-07-22

Applicant: UNIVERSITY OF UTAH RESEARCH FOUNDATION

Inventor： WEI, Ai-Ping ,  HERRON, James N.

IPC: C12Q1 ,  G01N33

CPC classification number: G01N33/582 ,  C12Q1/6818 ,  Y10S436/80 ,  Y10S436/805

Abstract: Fluorescent energy transfer dyes capable of moving between a more stacked configuration to exhibit fluorescent quenching and a more spaced configuration to exhibit fluorescence can be conjugated to a peptide epitope or nucleic acid for use in the detection of an unknown antibody in bulk solution. The resulting labelled peptide reagent can be used in an immunoassay procedure by placing it in bulk solution along with the unknown antibody to be detected. When the antibody binds to the peptide epitope, the pair of dyes carried by the peptide epitope will have their configuration altered from a stacked to an unstacked configuration and will exhibit a fluorescent increase in response thereto.