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公开(公告)号：EP2920199B1
公开(公告)日：2018-03-07
申请号：EP14832966.7
申请日：2014-05-07
申请人： Daewoong Co., Ltd.
CPC分类号： C12N9/52 , C07K14/33 , C12Y304/24069
摘要： A method of making botulinum toxin comprises treating a culture of a botulinum toxin-producing strain with acid to precipitate a botulinum toxin; adding buffer to the precipitated botulinum toxin, followed by treatment with a protease inhibitor and nuclease, thereby extracting the botulinum toxin; treating the extracted botulinum toxin with acid to precipitate the botulinum toxin and dissolving the precipitate in buffer; and purifying the botulinum toxin by anion exchange chromatography. The use of the method makes it possible to produce a high-purity botulinum toxin by a simple process. The botulinum toxin produced by the method has high purity, and thus has an increased ability to act in a local area. Thus, the systemic circulation of the botulinum toxin is reduced to increase the safety. Accordingly, the botulinum toxin can be used for treatment of neuromuscular disorders, removal of wrinkles, and treatment of spastic hemiplegia and cerebral palsy.
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公开(公告)号：EP2920199A1
公开(公告)日：2015-09-23
申请号：EP14832966.7
申请日：2014-05-07
申请人： Daewoong Co., Ltd.
CPC分类号： C12N9/52 , C07K14/33 , C12Y304/24069
摘要： The present invention relates to a method for production of a botulinum toxin, and more particularly to a method for preparation of botulinum toxin, the method comprising the steps of: (a) treating a culture of a botulinum toxin-producing strain with acid to precipitate a botulinum toxin; (b) adding buffer to the precipitated botulinum toxin, followed by treatment with a protease inhibitor and nuclease, thereby extracting the botulinum toxin; (c) treating the extracted botulinum toxin with acid to precipitate the botulinum toxin and dissolving the precipitate in buffer; and (d) purifying the botulinum toxin by anion exchange chromatography. The use of the method of the invention toxin makes it possible to produce a high-purity botulinum toxin by a simple process, suggesting that the method is very economical and efficient. The botulinum toxin produced by the method of the invention has high purity compared to botulinum toxins produced by conventional methods, and thus has an increased ability to act in a local area. Thus, the systemic circulation of the botulinum toxin, which can result in side effects, is reduced to increase the safety. Accordingly, the botulinum toxin of the invention can be used for various purposes, including treatment of neuromuscular disorders, removal of wrinkles, and treatment of spastic hemiplegia and cerebral palsy.
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公开(公告)号：EP3383889A1
公开(公告)日：2018-10-10
申请号：EP16870958.2
申请日：2016-11-23
申请人： DAEWOONG CO., LTD.
CPC分类号： C12N9/52 , B01D15/362 , B01D15/363 , B01D61/145 , B01D61/147 , B01D61/58 , B01D2315/16 , C07K14/33 , C12Y304/24069
摘要： The present invention relates to a method for producing a botulinum toxin comprising:(a) treating a culture of a botulinum toxin-producing strain with acid to form a botulinum toxin-containing precipitate; (b) adding a buffer to the botulinum toxin-containing precipitate of step (a), followed by clarification by at least one method selected from the group consisting of depth filtration (DF), microfiltration (MF), ultrafiltration (UF), sterile filtration, membrane chromatography (MC) and centrifugation; (c) subjecting the botulinum toxin-containing solution of step (b) to UF diafiltration, ammonium sulfate precipitation or hydrochloric acid precipitation, and then diluting a retentate resulting from the UF diafiltration in a buffer or dissolving a precipitate resulting from the ammonium sulfate precipitation or hydrochloric acid precipitate in a buffer; and (d) subjecting the retentate dilution, ammonium sulfate precipitate solution or hydrochloric acid precipitate solution of step (c) to anion-exchange chromatography (AEX) to purify the botulinum toxin.
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