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    Method for quantification of neurotoxin
    1.
    发明授权
    Method for quantification of neurotoxin 有权
    神经毒素定量方法

    公开(公告)号：US08949033B2

    公开(公告)日：2015-02-03

    申请号：US12298908

    申请日：2006-04-28

    申请人： Tetsuhiro Harakawa ,  Hirotoshi Nakano ,  Yasushi Torii ,  Sachio Okuda ,  Ryuji Kaji ,  Takashi Sakamoto ,  Motohide Takahashi ,  Setsuji Ishida

    发明人： Tetsuhiro Harakawa ,  Hirotoshi Nakano ,  Yasushi Torii ,  Sachio Okuda ,  Ryuji Kaji ,  Takashi Sakamoto ,  Motohide Takahashi ,  Setsuji Ishida

    IPC分类号： G01N33/50 ,  G01N33/94

    CPC分类号： G01N33/5088 ,  G01N33/94

    摘要： The present invention relates to a method for quantitatively measuring the muscular relaxing activity of a neurotoxin. Specifically, based on an extent of the activity of muscular relaxation of a neurotoxin from bacteria of Clostridium, the present invention relates to a method for quantification of the efficacy (potential and/or diffusion reaction) of a neurotoxin comprising the following steps of: (a) administering a neurotoxin to the hind leg muscle of one of hind legs of a non-human mammal; (b) applying electric stimulus to said non-human mammal; (c) measuring a compound muscle action potential (CMAP) by contraction of said hind leg muscle to which the neurotoxin is administered and/or of the hind leg muscle of the other hind leg to which the neurotoxin is not administered; and (d) taking amplitude data from the compound muscle action potential (CMAP) obtained by the measurement in step (c) and analyzing an extent of a decrease in amplitude to thereby quantify the efficacy of the muscular relaxing activity by the neurotoxin. In contrast to the mouse LD50 currently used as a potential unit of a botulinum toxin which is measurable at a level of only several units, the quantification method of the efficacy of a neurotoxin of the present invention allows for measurement at a level of as low as 0.01 to 1 unit and hence is a method with a high sensitivity, reproducibility and accuracy.

    摘要翻译： 本发明涉及一种定量测定神经毒素的肌肉松弛活性的方法。 具体而言，本发明是基于梭菌属细菌的神经毒素的肌肉弛豫活动的程度，本发明涉及一种定量神经毒素的功效（潜在和/或扩散反应）的方法，包括以下步骤：（ a）将神经毒素施用于非人哺乳动物后腿之一的后腿肌肉; （b）对所述非人哺乳动物施加电刺激; （c）通过神经毒素施用的所述后腿肌肉的收缩和/或不施用神经毒素的另一后腿的后腿肌肉测定复合肌肉动作电位（CMAP）; 和（d）从步骤（c）中测定得到的复合肌肉动作电位（CMAP）获取振幅数据，分析振幅减小的程度，由此量化神经毒素对肌肉松弛活性的功效。 与目前用作肉毒杆菌毒素的潜在单位的小鼠LD50相比，其可测量的水平仅为几个单位，本发明的神经毒素的功效的定量方法允许测量水平低至 0.01〜1单位，因此是具有高灵敏度，重现性和准确性的方法。

    Method for quantification of neurotoxin
    2.
    发明授权
    Method for quantification of neurotoxin 有权

    公开(公告)号：US08577618B2

    公开(公告)日：2013-11-05

    申请号：US12298908

    申请日：2006-04-28

    申请人： Tetsuhiro Harakawa ,  Hirotoshi Nakano ,  Yasushi Torii ,  Sachio Okuda ,  Ryuji Kaji ,  Takashi Sakamoto ,  Motohide Takahashi ,  Setsuji Ishida

    发明人： Tetsuhiro Harakawa ,  Hirotoshi Nakano ,  Yasushi Torii ,  Sachio Okuda ,  Ryuji Kaji ,  Takashi Sakamoto ,  Motohide Takahashi ,  Setsuji Ishida

    IPC分类号： G01N33/50

    摘要： The present invention relates to a method for quantitatively measuring the muscular relaxing activity of a neurotoxin. Specifically, based on an extent of the activity of muscular relaxation of a neurotoxin from bacteria of Clostridium, the present invention relates to a method for quantification of the efficacy (potential and/or diffusion reaction) of a neurotoxin comprising the following steps of: (a) administering a neurotoxin to the hind leg muscle of one of hind legs of a non-human mammal; (b) applying electric stimulus to said non-human mammal; (c) measuring a compound muscle action potential (CMAP) by contraction of said hind leg muscle to which the neurotoxin is administered and/or of the hind leg muscle of the other hind leg to which the neurotoxin is not administered; and (d) taking amplitude data from the compound muscle action potential (CMAP) obtained by the measurement in step (c) and analyzing an extent of a decrease in amplitude to thereby quantify the efficacy of the muscular relaxing activity by the neurotoxin. In contrast to the mouse LD50 currently used as a potential unit of a botulinum toxin which is measurable at a level of only several units, the quantification method of the efficacy of a neurotoxin of the present invention allows for measurement at a level of as low as 0.01 to 1 unit and hence is a method with a high sensitivity, reproducibility and accuracy.

    METHOD FOR QUANTIFICATION OF TITER OF NEUTRALIZING ANTIBODY TO NEUROTOXIN
    6.
    发明申请
    METHOD FOR QUANTIFICATION OF TITER OF NEUTRALIZING ANTIBODY TO NEUROTOXIN 审中-公开
    中性抗体对神经毒素的定量的方法

    公开(公告)号：US20110105940A1

    公开(公告)日：2011-05-05

    申请号：US12673980

    申请日：2008-08-20

    申请人： Yasushi Torii ,  Tetsuhiro Harakawa ,  Yoshitaka Goto ,  Akihiro Ginnaga ,  Ryuji Kaji ,  Shunji Kozaki ,  Motohide Takahashi

    发明人： Yasushi Torii ,  Tetsuhiro Harakawa ,  Yoshitaka Goto ,  Akihiro Ginnaga ,  Ryuji Kaji ,  Shunji Kozaki ,  Motohide Takahashi

    IPC分类号： A61B5/0488

    CPC分类号： G01N33/6854 ,  A61B5/0488 ,  G01N2333/33 ,  G01N2469/20

    摘要： A method for quantification of a titer of a neutralizing antibody to a neurotoxin is disclosed. The method comprises the steps: (a) mixing a standard sample containing a fixed amount of a neurotoxin and a test sample containing a neutralizing antibody to said neurotoxin; (b) administering the mixture obtained in step (a) into the muscle of a non-human mammal; (c) applying electric stimulus to said non-human mammal; (d) measuring a compound muscle action potential (CMAP) due to contraction of the muscle of said mammal by application of electric stimulus with an electromyograph; and (e) analyzing CMAP amplitude data obtained in step (d) for an extent of decrease in amplitude by a non-neutralized neurotoxin to thereby quantify a titer of the neutralizing antibody contained in the test sample.

    摘要翻译： 公开了定量神经毒素中和抗体滴度的方法。 该方法包括以下步骤：（a）将含有固定量的神经毒素的标准样品和含有中和抗体的测试样品与所述神经毒素混合; （b）将步骤（a）中获得的混合物给予非人哺乳动物的肌肉; （c）对所述非人哺乳动物施加电刺激; （d）通过用肌电图仪施加电刺激来测量由于所述哺乳动物的肌肉收缩引起的复合肌肉动作电位（CMAP）; 和（e）通过未中和的神经毒素分析在步骤（d）中获得的CMAP振幅数据的幅度减小，从而定量测试样品中所含的中和抗体的滴度。