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    METHOD FOR QUANTIFICATION OF TITER OF NEUROTOXIN-NEUTRALIZING ANTIBODY
    1.
    发明公开
    METHOD FOR QUANTIFICATION OF TITER OF NEUROTOXIN-NEUTRALIZING ANTIBODY 有权
    定量测定神经毒素中和抗体滴度的方法

    公开(公告)号：EP2189790A1

    公开(公告)日：2010-05-26

    申请号：EP08827849.4

    申请日：2008-08-20

    申请人： Juridical Foundation The Chemo-Sero-Therapeutic Research Institute ,  JAPAN as represented by DIRECTOR GENERAL OF NATIONAL INSTITUTE OF INFECTIOUS DISEASES

    发明人： TORII, Yasushi ,  HARAKAWA, Tetsuhiro ,  GOTO, Yoshitaka ,  GINNAGA, Akihiro ,  KAJI, Ryuji ,  KOZAKI, Shunji ,  TAKAHASHI, Motohide

    IPC分类号： G01N33/48 ,  G01N33/15

    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： 公开了定量测定神经毒素中和抗体效价的方法。 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混合到所述神经毒素中; （b）将步骤（a）中获得的混合物给予非人类哺乳动物的肌肉; （c）对所述非人类哺乳动物施加电刺激; （d）通过用肌电图应用电刺激测量所述哺乳动物肌肉收缩引起的复合肌肉动作电位（CMAP）; 和（e）分析步骤（d）中获得的CMAP幅度数据对于未中和的神经毒素的幅度降低程度，从而定量测试样品中含有的中和抗体的效价。

    METHOD OF QUANTIFYING NEUROTOXIN
    2.
    发明授权
    METHOD OF QUANTIFYING NEUROTOXIN 无效
    方法定量NEURO毒素

    公开(公告)号：EP2015065B1

    公开(公告)日：2011-10-05

    申请号：EP06745899.2

    申请日：2006-04-28

    申请人： Juridical Foundation The Chemo-Sero-Therapeutic Research Institute ,  JAPAN as represented by DIRECTOR GENERAL OF NATIONAL INSTITUTE OF INFECTIOUS DISEASES

    发明人： HARAKAWA, Tetsuhiro ,  NAKANO, Hirotoshi ,  TORII, Yasushi ,  OKUDA, Sachio ,  KAJI, Ryuji ,  SAKAMOTO, Takashi ,  TAKAHASHI, Motohide ,  ISHIDA, Setsuji

    IPC分类号： G01N33/15 ,  G01N33/50

    CPC分类号： G01N33/5088 ,  G01N33/94

    摘要： A method of quantitatively measuring the muscle relaxant action of a neurotoxin. More specifically speaking, a method of quantifying the efficacy (titer and/or diffusion reaction) of a neurotoxin based on the degree of the muscle relaxant action of a neurotoxin originating in a bacterium belonging to the genus Clostridium, which is characterized by comprising: (a) administering the neurotoxin to one hinder leg of a nonhuman mammal; (b) electrically stimulating the nonhuman mammal; (c) measuring the compound muscle action potential (CMAP) caused by the contraction of muscles in the hinder leg to which the neurotoxin has been administered and/or muscles in the other hinder leg to which the neurotoxin has not been administered by using an electromyograph; and (d) taking out the amplitude data from the compound muscle action potential (CMAP) obtained by the measurement in the step (c) and analyzing the degree of a decrease in the amplitude to thereby quantify the efficacy of the muscle relaxant action by the neurotoxin. Although mouse LD50 that has been employed as a titer unit of the botulin toxin allows the measurement only at a several unit level, the above-described method of quantifying the efficacy of a neurotoxin enables the measurement at 0.01 to 1 unit level. That is, this method is excellent in sensitivity, reproducibility and accuracy.

    METHOD OF QUANTIFYING NEUROTOXIN
    5.
    发明公开
    METHOD OF QUANTIFYING NEUROTOXIN 无效
    量化神经毒素的方法

    公开(公告)号：EP2015065A1

    公开(公告)日：2009-01-14

    申请号：EP06745899.2

    申请日：2006-04-28

    申请人： Juridical Foundation The Chemo-Sero-Therapeutic Research Institute ,  Japan as Repres. by Director Gral of National Institute of Infectious Diseases

    发明人： HARAKAWA, Tetsuhiro ,  NAKANO, Hirotoshi ,  TORII, Yasushi ,  OKUDA, Sachio ,  KAJI, Ryuji ,  SAKAMOTO, Takashi ,  TAKAHASHI, Motohide ,  ISHIDA, Setsuji

    IPC分类号： G01N33/15 ,  G01N33/50

    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 LD 50 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： 本发明涉及定量测定神经毒素的肌肉松弛活性的方法。 具体而言，基于来自梭菌细菌的神经毒素的肌肉松弛活性的程度，本发明涉及定量神经毒素的功效（潜在和/或扩散反应）的方法，其包括以下步骤:( a）将神经毒素给予非人类哺乳动物后腿之一的后腿肌肉; （b）向所述非人类哺乳动物施加电刺激; （c）通过所述神经毒素给予的所述后腿肌肉和/或未给予所述神经毒素的另一后腿的后腿肌肉的收缩来测量复合肌肉动作电位（CMAP）; （d）从通过步骤（c）中的测量获得的复合肌肉动作电位（CMAP）获取振幅数据并分析振幅降低的程度，从而量化神经毒素对肌肉放松活动的功效。 与目前用作肉毒毒素潜在单位的小鼠LD50相比，所述肉毒杆菌毒素可以以仅几个单位的水平测量，本发明神经毒素功效的定量方法允许测量的水平低至 0.01到1单位，因此是一种高灵敏度，重现性和准确性的方法。

    REMEDY FOR HYPERMYOTONIA
    6.
    发明公开
    REMEDY FOR HYPERMYOTONIA 审中-公开
    MITTEL ZUR BEHANDLUNG VON HYPERMYOTONIE

    公开(公告)号：EP1491205A1

    公开(公告)日：2004-12-29

    申请号：EP03715663.5

    申请日：2003-03-31

    申请人： Juridical Foundation, The Chemo-Sero-Therapeutic Research Institute

    发明人： KAJI, Ryuji ,  KOZAKI, Shunji ,  OGUMA, Keiji

    IPC分类号： A61K38/00 ,  A61K45/00 ,  A61K47/42 ,  A61P21/02 ,  A61P25/00 ,  A61P25/14 ,  A61P27/02 ,  A61P43/00

    CPC分类号： A61K38/4886

    摘要： A remedy for muscle hyperactivity, comprising a purified botulinum neurotoxin as an active ingredient.

    摘要翻译： 用于治疗高水平症的药物包括纯化的肉毒神经毒素。 活动性：肌肉活跃。 肉毒杆菌神经毒素从培养的B型微生物培养物获得，并在离子交换柱上分离，然后用β-乳糖凝胶柱分离。 通过胰蛋白酶和β-乳糖凝胶亲和层析纯化肉毒神经毒素L。 将获得的肉毒神经毒素L透析至pH8.0,10mM磷酸盐缓冲液，收集在β-乳糖凝胶亲和层析柱上不吸收的级分，得到肉毒神经毒素B.其活性通过ip注射到小鼠中进行测定。 用20mM磷酸钠缓冲液将pH调节至6.0,7.0或8.0，浓度为500MLD / 0.25ml（MLD是小鼠腹腔注射时的最小致死剂量）。 将不同的pH样品与等量的1mg / ml的人血清白蛋白（HSA）混合，并在4℃冷藏并在-80℃下冷冻。 对于具有HSA的样品，90天后的活性在500MLD时不变，而在未添加HSA的样品下，1MLD或更低。 行动机制：源材料无。