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Assay for quantifying clostridial neurotoxin

A technology for neurotoxins and clostridial toxins, applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of lack of regulatory accuracy in quantitative methods, untimely mouse killing tests, etc.

Inactive Publication Date: 2012-08-15
MERZ PHARMA GMBH & CO KGAA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0015] However, the quantification methods of the prior art cited above lack the precision required for regulatory agency certification
Therefore, those published methods cannot be used for administrative purposes, and instead require an untimely mouse kill test

Method used

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  • Assay for quantifying clostridial neurotoxin
  • Assay for quantifying clostridial neurotoxin
  • Assay for quantifying clostridial neurotoxin

Examples

Experimental program
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Embodiment 1

[0636]Mouse hemidiaphragms were prepared and added to organ baths containing Earle's Balanced Salt Solution for standard assays. A platinum electrode is installed on the phrenic nerve of the hemidiaphragm, and the nerve is electrically stimulated through the platinum electrode, which subsequently affects the contraction of the hemidiaphragm. Clamp the hemidiaphragm in an organ bath. During clamping, the stimulation is turned off, but turned on immediately after clamping. The current intensity of the stimulation is chosen so that the contractility of the diaphragm can be measured. After sustained contraction force can be determined, the medium is changed to medium containing botulinum neurotoxin. The time required to reach half the contraction force (paralysis time) was determined for each concentration (at least several times for each concentration) and plotted against the concentration of botulinum neurotoxin added in the organ bath.

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Abstract

Method of measuring an effect induced to a muscle tissue by a clostridial neurotoxin, comprising: (a) contacting a muscle tissue or a cell culture with a sample comprising said clostridial neurotoxin; (c) measuring said effect induced to said muscle tissue by said clostridial neurotoxin; wherein step (c) is performed in the absence of said sample.

Description

technical field [0001] The present invention relates to a method for in vitro determination of an unknown concentration of a Clostridial neurotoxin in a sample with reference to a known concentration of the Clostridial toxin in a reference sample. The method may include electrically stimulating muscle tissue that has been in contact with said sample, and comparing the respective effects induced on said muscle tissue, thereby determining said unknown concentration. This method can also be used to assess the relative potency of Clostridial neurotoxins in a sample relative to a reference standard. Background technique [0002] In recent years, botulinum neurotoxin has become the standard formulation for the treatment of localized dystonic and spastic indications. Pharmaceutical formulations are commercially available, for example from Ipsen Ltd. or Allergan Inc. Highly pure neurotoxins free of any other Clostridial proteins are available from, for example, Merz Pharmaceuti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61B5/0488A61B5/11G01N33/50G01N33/94
CPCG01N33/5088G01N33/56911G01N33/94G01N2333/33
Inventor 杰德·J·曼德哈罗德·泰勒马丁·韦卡尔·海因茨·艾斯勒
Owner MERZ PHARMA GMBH & CO KGAA