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Method of producing clostridium botulinum toxin using media containing plant-derived components and flexible closed container

一种肉毒梭菌、密闭容器的技术，应用在生物化学设备和方法、酵素、细菌等方向，能够解决存在高危因素、高投资成本、大劳动力等问题

Inactive Publication Date: 2011-06-08
MEDY TOX INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This system requires a lot of labor and high investment costs, and there are high risk factors in this system in order to meet the current good manufacturing practice (cGMP, current good manufacturing practice) requirements

Method used

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  • Method of producing clostridium botulinum toxin using media containing plant-derived components and flexible closed container
  • Method of producing clostridium botulinum toxin using media containing plant-derived components and flexible closed container

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0049] Example 1: Validation of medium composition to produce high levels of Clostridium botulinum toxin type A expressed outside the cell

[0050] The fact that a large amount of botulinum toxin type A is expressed extracellularly in media free of animal-derived components was verified using various plant media.

[0051] The media used are shown in Table 1 below.

[0052]

[0053] [Table 1]

[0054] [sheet]

[0055]

[0056] In Table 1, products manufactured by Beckton Dickinson and Fluka were used as soytone and phytotryptone, respectively. In addition, products manufactured by Becton-Dickinson were used as plant peptone and yeast extract, and products manufactured by Merck were used as glucose. The medium components of Table 1 were dissolved in water for injection (WFI), and the pH of the resulting solution was adjusted to 7.2 using NaOH.

[0057] In addition, 1% by weight of N-Z amine A (Sigma (Sigma): animal origin), 2% by weight of Bacto peptone (Proteose Pep...

example 2

[0075] In addition to not using Cultibag while culturing TM This example was performed in the same manner as Example 1 except that shaking was performed.

[0076] As a result, there was no substantial difference between the amount of toxin produced and that of Example 1.

example 3

[0077] Example 3: Isolation of meat type A from culture supernatant using media containing plant-derived components Clostridium toxin

[0078] Clostridium botulinum was cultured using medium 5 of Example 1 or 2, and botulinum toxin type A was isolated from the supernatant from which the cells had been removed. The culture conditions and time and the cell removal process were the same as in Example 1.

[0079] 0.1 N sulfuric acid was added to the bag containing 20 L of the culture supernatant to adjust the pH of the resulting solution to be in the range of 3.0 to 4.5, and thus, precipitate the toxin protein. Perform this procedure in a disposable sample bag that does not come into contact with the outside.

[0080]Next, when the precipitation was completed, the resulting solution was filtered through a pre-sterilized disposable filtration system (disposable sheet 2000.2 μm filter, manufactured by Sartorius Corporation) that was not in contact with the outside to remove the...

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PUM

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Abstract

Provided are a method of producing Clostridium botulinum toxin by using a media containing plant-derived components, and a method of producing Clostridium botulinum toxin by using a flexible closed container.

Description

technical field [0001] This application claims the benefit of Korean Patent Application No. 10-2008-0046152 filed with the Korean Intellectual Property Office on May 19, 2008, the disclosure of which is incorporated herein by reference in its entirety. [0002] The present invention relates to a method for the preparation of Clostridium botulinum toxin using a culture medium containing plant-derived components and a flexible, closed container. Background technique [0003] From 1890 to the present, Clostridium (Clostridium) bacterial strains capable of producing neurotoxic toxins have been found, and the research on the properties of toxins produced by Clostridium bacterial strains has been carried out for 70 years (Stanzi (Schant, E.J.) et al., Microbiol. Rev., Vol. 56; 80; 1992). [0004] The genus Clostridium has more than 127 species and is grouped according to their morphology and function. The anaerobic Gram-positive bacterium Clostridium botulinum produces polypepti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/04
CPCC12N9/52C12Y304/24069C12N1/20C12P21/02C07K14/33
Inventor 丁贤壕梁起赫金学佑李炳国尹英淑洪衡杓
Owner MEDY TOX INC
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