Protocol and software for multiplex real-time PCR quantification based on the different melting temperatures of amplicons
    3.
    发明申请
    Protocol and software for multiplex real-time PCR quantification based on the different melting temperatures of amplicons 审中-公开
    基于扩增子的不同融合温度的多重实时PCR定量方法和软件

    公开(公告)号:US20050053950A1

    公开(公告)日:2005-03-10

    申请号:US10658602

    申请日:2003-09-08

    IPC分类号: B01L7/00 C12P19/34 C12Q1/68

    摘要: The present invention provides a new protocol for quantifying multiplex real-time polymerase chain reaction (PCR). In particular, the present invention provides methods of quantifying multiple PCR products or amplicons in a single real-time PCR reaction based on the different melting temperatures (Tm) of each amplicon and the emission changes of double stranded DNA dyes such as SYBR Green I when amplicons are in duplex or in separation. For a specific amplicon with a Tm, the emission difference between the emission reading taken at a temperature below the Tm and the emission reading taken at a temperature above the Tm corresponds to the emission value of the amplicon in duplex. Accordingly, the emission difference of each amplicon in a single PCR reaction can be used to quantify each amplicon. The present invention further provides computer programs or computer products which perform the methods described herein.

    摘要翻译: 本发明提供了一种用于量化多重实时聚合酶链反应(PCR)的新方案。 特别地,本发明提供了基于每个扩增子的不同解链温度(Tm)和双链DNA染料如SYBR Green I的发射变化在单次实时PCR反应中定量多个PCR产物或扩增子的方法, 扩增子是双链体或分离的。 对于具有Tm的特异性扩增子,在低于Tm的温度下获取的发射读数与在高于Tm的温度下获得的发射读数之间的发射差异对应于双链体扩增子的发射值。 因此,单个PCR反应中每个扩增子的发射差异可用于量化每个扩增子。 本发明还提供了执行本文描述的方法的计算机程序或计算机产品。

    MULTIPLEX ASSAY METHOD FOR MIXED CELL POPULATIONS
    5.
    发明申请
    MULTIPLEX ASSAY METHOD FOR MIXED CELL POPULATIONS 审中-公开
    用于混合细胞群体的多重测定方法

    公开(公告)号:US20090088341A1

    公开(公告)日:2009-04-02

    申请号:US12060752

    申请日:2008-04-01

    IPC分类号: C40B30/10

    CPC分类号: G01N33/5005

    摘要: Disclosed is an in vitro assay for evaluating cellular activity, in which a culture is provided that contains one or more histologically different isolated mammalian cell-line that stably and constitutively express fluorescent proteins having a different emission spectra. The culture is assessed for cellular activity by quantifying fluorescence or detecting a pattern of fluorescence from fluorescent proteins present in the culture. In some embodiments, the cellular activity of the cell-lines is assessed by determining one or more of the growth rate, migration potential, or tubule formation potential of the cell-lines using the quantified fluorescence or pattern of fluorescence. Also disclosed are cell-lines have been stably transfected with mammalian expression plasmids that constitutively express different fluorescent proteins. Kits for performing the disclosed assays, which include the disclosed fluorescent cell-lines, are also disclosed.

    摘要翻译: 公开了用于评估细胞活性的体外测定法,其中提供了含有一种或多种组织学上不同的分离的哺乳动物细胞系的培养物,其稳定和组成型表达具有不同发射光谱的荧光蛋白质。 通过定量荧光或从存在于培养物中的荧光蛋白检测荧光图案来评估培养物的细胞活性。 在一些实施方案中,通过使用定量的荧光或荧光图案确定细胞系的生长速率,迁移电位或小管形成潜能来评估细胞系的细胞活性。 还公开了细胞系已经用组成型表达不同荧光蛋白的哺乳动物表达质粒稳定转染。 还公开了用于进行所公开的测定的试剂盒,其包括所公开的荧光细胞系。

    Functional role of adrenomedullin (AM) and the gene related product (PAMP) in human pathology and physiology
    7.
    发明授权
    Functional role of adrenomedullin (AM) and the gene related product (PAMP) in human pathology and physiology 有权
    肾上腺髓质素(AM)和基因相关产物(PAMP)在人类病理和生理学中的功能作用

    公开(公告)号:US07939639B2

    公开(公告)日:2011-05-10

    申请号:US12569821

    申请日:2009-09-29

    摘要: The methods of the present invention demonstrate that adrenomedullin (AM) is expressed in human cancer cell lines of diverse origin and functions as a universal autocrine growth factor driving neoplastic proliferation. The present invention provides for AM peptides and AM antibodies useful in therapeutic, pharmacologic and physiologic compositions. The present invention additionally provides for methods of diagnosis, treatment and prevention of disease utilizing compositions comprising the AM peptides and antibodies of the present invention. The methods of the present invention also provide for experimental models for use in identifying the role of AM in pancreatic physiology. The methods pertaining to rat isolated islets have show that AM inhibits insulin secretion in a dose-dependent manner. The monoclonal antibody MoAb-G6, which neutralizes AM bioactivity, was show by the methods of the present invention to increase insulin release fivefold, an effect that was reversed by the addition of synthetic AM.

    摘要翻译: 本发明的方法证明,肾上腺髓质素(AM)在不同来源的人类癌细胞系中表达,并且作为驱动肿瘤增殖的通用自分泌生长因子起作用。 本发明提供可用于治疗,药理和生理组合物的AM肽和AM抗体。 本发明另外提供了使用包含本发明的AM肽和抗体的组合物来诊断,治疗和预防疾病的方法。 本发明的方法还提供了用于鉴定AM在胰腺生理学中的作用的实验模型。 与大鼠分离的胰岛有关的方法表明,AM以剂量依赖的方式抑制胰岛素分泌。 通过本发明的方法显示了中和AM生物活性的单克隆抗体MoAb-G6,以增加胰岛素释放的五倍,这是通过加入合成AM而逆转的作用。

    Determination of AM-binding proteins and the association of adrenomedullin (AM) therewith
    8.
    发明授权
    Determination of AM-binding proteins and the association of adrenomedullin (AM) therewith 有权
    AM结合蛋白的测定和肾上腺髓质素(AM)与其结合

    公开(公告)号:US07659081B2

    公开(公告)日:2010-02-09

    申请号:US12236418

    申请日:2008-09-23

    IPC分类号: G01N33/53

    摘要: The present invention provides methods for the isolation, identification, and purification of adrenomedullin (AM)-binding proteins. Also, provided are methods for utilizing the purified AM-binding proteins, or functional portions thereof, to diagnose, treat, and monitor AM-related diseases, for example, diseases or disorders associated with abnormally elevated AM levels. In addition, the present invention provides a newly identified complex between AM and a specific AM-binding protein 1 (AMBP-1); which has been isolated and identified herein as factor H (fH). The invention also provides AM/AMBP complexes, particularly AM/FH complexes, and antibodies specifically reactive with this complexes. Further provided are methods for identifying and purifying complexes of AM and an AM binding protein using anti-AM/fH antibodies, and methods for treating conditions such as cancer or diabetes utilizing compositions comprising these antibodies. The present invention additionally provides methods for identifying antagonists agents that inhibit the function of AM, factor H, or the AM/factor H complex. The invention also provides methods for treating conditions such as cancer or diabetes using these antagonist agents.

    摘要翻译: 本发明提供了分离,鉴定和纯化肾上腺髓质素(AM)结合蛋白的方法。 此外,提供了利用纯化的AM结合蛋白或其功能部分来诊断,治疗和监测AM相关疾病的方法,例如与异常升高的AM水平相关的疾病或病症。 此外,本发明提供了AM和特异性AM结合蛋白1(AMBP-1)之间新鉴定的复合物; 其在本文中被分离和鉴定为因子H(fH)。 本发明还提供AM / AMBP复合物,特别是AM / FH复合物和与这些复合物特异性反应的抗体。 进一步提供了使用抗AM / fH抗体鉴定和纯化AM和AM结合蛋白的复合物的方法,以及使用包含这些抗体的组合物治疗诸如癌症或糖尿病的病症的方法。 本发明另外提供用于鉴定抑制AM,因子H或AM /因子H复合物的功能的拮抗剂的方法。 本发明还提供了使用这些拮抗剂治疗诸如癌症或糖尿病的病症的方法。

    Functional role of adrenomedullin (AM) and the gene-related product (PAMP) in human pathology and physiology
    9.
    发明授权
    Functional role of adrenomedullin (AM) and the gene-related product (PAMP) in human pathology and physiology 失效
    肾上腺髓质素(AM)和基因相关产物(PAMP)在人类病理和生理学中的功能作用

    公开(公告)号:US07622272B2

    公开(公告)日:2009-11-24

    申请号:US11517599

    申请日:2006-09-05

    摘要: The methods of the present invention demonstrate that adrenomedullin (AM) is expressed in human cancer cell lines of diverse origin and functions as a universal autocrine growth factor driving neoplastic proliferation. The present invention provides for AM peptides and AM antibodies useful in therapeutic, pharmacologic and physiologic compositions. The present invention additionally provides for methods of diagnosis, treatment and prevention of disease utilizing compositions comprising the AM peptides and antibodies of the present invention. The methods of the present invention also provide for experimental models for use in identifying the role of AM in pancreatic physiology. The methods pertaining to rat isolated islets have show that AM inhibits insulin secretion in a dose-dependent manner. The monoclonal antibody MoAb-G6, which neutralizes AM bioactivity, was show by the methods of the present invention to increase insulin release fivefold, an effect that was reversed by the addition of synthetic AM.

    摘要翻译: 本发明的方法证明,肾上腺髓质素(AM)在不同来源的人类癌细胞系中表达,并且作为驱动肿瘤增殖的通用自分泌生长因子起作用。 本发明提供可用于治疗,药理和生理组合物的AM肽和AM抗体。 本发明另外提供了使用包含本发明的AM肽和抗体的组合物来诊断,治疗和预防疾病的方法。 本发明的方法还提供了用于鉴定AM在胰腺生理学中的作用的实验模型。 与大鼠分离的胰岛有关的方法表明,AM以剂量依赖的方式抑制胰岛素分泌。 通过本发明的方法显示了中和AM生物活性的单克隆抗体MoAb-G6,以增加胰岛素释放的五倍,这是通过加入合成AM而逆转的作用。