Method for producing high-purity soluble thrombomodulin

C - Chemistry – Metallurgy – 07 – K

Patent

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C07K 1/16 (2006.01) A61P 7/02 (2006.01) C12N 15/09 (2006.01) C12P 21/02 (2006.01) A61K 38/16 (2006.01)

Patent

CA 2681294

It is intended to obtain soluble thrombomodulin which substantially contains no soluble thrombomodulin variant that can be formed under an acidic condition. According to the invention, a method for producing soluble thrombomodulin which substantially contains no soluble thrombomodulin variant from a soluble thrombomodulin-containing material which contains or is suspected to contain a soluble thrombomodulin variant that can be formed under an acidic condition, comprising the steps of: subjecting the soluble thrombomodulin-containing material to an anion exchanger or hydroxyapatite; and obtaining an eluted fraction containing soluble thrombomodulin which substantially contains no soluble thrombomodulin variant by applying a linear or stepwise gradient of salt concentration or a combination of linear and stepwise gradients thereof under a separation condition capable of separating the soluble thrombomodulin variant and performing identification in advance or while performing identification is provided.

L'invention vise à obtenir une thrombomoduline soluble qui ne contient pratiquement pas de variant de thrombomoduline soluble qui peut être formé dans une condition acide. Selon l'invention, le procédé de fabrication de thrombomoduline soluble qui ne contient pratiquement pas de variant de thrombomoduline soluble à partir d'une matière contenant une thrombomoduline soluble qui contient ou est supposée contenir un variant de thrombomoduline soluble qui peut être formé dans une condition acide, comprend les étapes consistant à : soumettre la matière contenant de la thrombomoduline soluble à un échangeur d'anions ou une hydroxyapatite ; et obtenir une fraction éluée contenant une thrombomoduline soluble qui ne contient pratiquement pas de variant de thrombomoduline soluble par application d'un gradient linéaire ou par étapes de concentration de sel ou une combinaison de gradients linéaire ou par étapes de celui-ci dans une condition de séparation capable de séparer le variant de thrombomoduline soluble et d'effectuer une identification à l'avance ou tout en effectuant une identification.

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