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EP-3269725-B1 - METHOD FOR PRODUCING PROTEIN COMPOSITION, AND PROTEIN COMPOSITION

EP3269725B1EP 3269725 B1EP3269725 B1EP 3269725B1EP-3269725-B1

Inventors

  • SOMAMOTO, Satoshi
  • KAWABATA, SHINGO

Dates

Publication Date
20260513
Application Date
20160309

Claims (4)

  1. A method for producing a protein composition containing a protein (A), a radical scavenger (RS), and at least one hydrogen-bond-formable compound (HC) consisting of amino acids, the method comprising: a sterilization step of radiosterilizing an unsterilized protein composition, wherein the unsterilized protein composition contains the protein (A), the radical scavenger (RS), and the hydrogen-bond-formable compound (HC), the protein (A) contains at least one functional group selected from the group consisting of sulfide, amide, hydroxyl, amino, and carboxyl groups, the at least one functional group in the protein (A) is capable of binding to the at least one functional group in the hydrogen-bond-formable compound (HC) via a hydrogen bond, the unsterilized protein composition has a water content of 0.01 to 30% by weight based on the weight of the unsterilized protein composition, a weight ratio of the radical scavenger (RS) to the protein (A) in the unsterilized protein composition, being the radical scavenger (RS)/protein (A), is 0.01 to 1.0, a molar ratio of a total molar number of functional groups in the hydrogen-bond-formable compound (HC) to a total molar number of functional groups in the protein (A) in the unsterilized protein composition, being the total molar number of functional groups in the hydrogen-bond-formable compound (HC)/the total molar number of functional groups in the protein (A), is 0.01 to 0.50, the hydrogen-bond-formable compound (HC) is at least one selected from the group consisting of tryptophan, tyrosine and histidine and the protein (A) is at least one selected from group consisting of SELP 8K, ProNectin F, ProNectin L, HRP-conjugated rabbit antibody, glucose oxidase and bovine serum albumin.
  2. The method for producing a protein composition according to claim 1, wherein the radical scavenger (RS) contains at least one selected from the group consisting of ascorbic acid, edaravone, vanillin, catechin, gallic acid, glutathione and chlorogenic acid.
  3. The method for producing a protein composition according to claim 1 or 2, wherein the radical scavenger (RS) has a radical scavenging ability against diphenylpicrylhydrazyl radicals of 0.01 to 90 mg Trolox eq/mg.
  4. A protein composition, comprising: a protein (A), wherein the protein composition further contains a radical scavenger (RS), and at least one hydrogen-bond-formable compound (HC) consisting of amino acids, the protein (A) contains at least one functional group selected from the group consisting of sulfide, amide, hydroxyl, amino, and carboxyl groups, the at least one functional group in the protein (A) is capable of binding to the at least one functional group in the hydrogen-bond-formable compound (HC) via a hydrogen bond, a weight ratio of the radical scavenger (RS) to the protein (A) in the protein composition, being radical scavenger (RS)/protein (A), is 0.01 to 1.0, a molar ratio of a total molar number of functional groups in the hydrogen-bond-formable compound (HC) to a total molar number of functional groups in the protein (A) in the protein composition, being the total molar number of functional groups in the hydrogen-bond-formable compound (HC)/the total molar number of functional groups in the protein (A), is 0.01 to 0.50, and the protein composition has been radiosterilized, the hydrogen-bond-formable compound (HC) is at least one selected from the group consisting of tryptophan, tyrosine and histidine and the protein (A) is at least one selected from group consisting of SELP 8K, ProNectin F, ProNectin L, HRP-conjugated rabbit antibody, glucose oxidase and bovine serum albumin.

Description

TECHNICAL FIELD The present invention relates to a method for producing a protein composition and a protein composition. BACKGROUND ART Proteins for use in medical applications and biochemical applications need to be sterilized before use. Sterilization is achieved by ethylene oxide gas sterilization, filtration sterilization, or radiosterilization. Radiosterilization is advantageous in terms of the absence of toxic residues (ethylene oxide gas) after sterilization as well as cost and validation. Radiosterilization of proteins unfortunately causes changes such as degradation and denaturation. Such changes in proteins are induced by reactions of active radicals (hydroxy radicals, oxygen radicals) generated by radiation with the proteins. Conventionally, the following methods have been employed in order to suppress changes in proteins induced by active radicals: a method in which proteins are irradiated with radiation under cooling conditions; a method in which water is removed from a radiation irradiation target; and a method in which a radical scavenger against active radicals is added (Patent Literatures 1 and 2); and a method of ionizing a protein, comprises exposing to ionizing radiation an at least substantially dry composition comprises a protein and a protective compound or combination of protective compounds (Patent Literature 3). Yet, the method in which proteins are irradiated with radiation under cooling conditions is not very effective, and the production cost for cooling is high. In addition, when sterilizing a highly hydrophilic protein or a protein that undergoes denaturation by complete removal of water, it is difficult to remove water from a radiation irradiation target. Further, in the case where a radical scavenger is used to prevent changes in proteins induced by active radicals, the radical scavenger needs to be added in a large amount, which unfortunately impairs physiological and physicochemical functions of the proteins. CITATION LIST - Patent Literature Patent Literature 1: JP-T 2003-527210Patent Literature 2: JP-T 2010-514747Patent Literature 3: WO 2008/081166 A1 SUMMARY OF INVENTION - Technical Problem The present invention aims to provide a method for producing a protein composition, the method being capable of suppressing changes such as degradation and denaturation of a protein which occur when a highly hydrophilic protein or a protein that undergoes denaturation by complete removal of water is irradiated with radiation. - Solution to Problem As a result of extensive studies, the present inventors arrived at the present invention. Specifically, the present invention is directed to a method for producing a protein composition containing a protein (A), a radical scavenger (RS), and at least one hydrogen-bond-formable compound (HC) consisting of amino acids, the method including a sterilization step of radiosterilizing an unsterilized protein composition, wherein the unsterilized protein composition contains the protein (A), the radical scavenger (RS), and the hydrogen-bond-formable compound (HC), the protein (A) contains at least one functional group selected from the group consisting of sulfide, amide, hydroxyl, amino, and carboxyl groups, the at least one functional group in the protein (A) is capable of binding to the at least one functional group in the hydrogen-bond-formable compound (HC) via a hydrogen bond, the unsterilized protein composition has a water content of 0.01 to 30% by weight based on the weight of the unsterilized protein composition, a weight ratio of the radical scavenger (RS) to the protein (A) in the unsterilized protein composition [radical scavenger (RS)/protein (A)] is 0.01 to 1.0, and a molar ratio of a total molar number of functional groups in the hydrogen-bond-formable compound (HC) to a total molar number of functional groups in the protein (A) in the unsterilized protein composition [the total molar number of functional groups in the hydrogen-bond-formable compound (HC)/the total molar number of functional groups in the protein (A)] is 0.01 to 0.50, the hydrogen-bond-formable compound (HC) is at least one selected from the group consisting of tryptophan, tyrosine and histidine, and the protein (A) is at least one selected from group consisting of SELP 8K, ProNectin F, ProNectin L, HRP-conjugated rabbit antibody, glucose oxidase and bovine serum albumin. The present invention also relates to a protein composition containing a protein (A), wherein the protein composition further contains a radical scavenger (RS), and at least one hydrogen-bond-formable compound (HC) consisting of amino acids, the protein (A) contains at least one functional group selected from the group consisting of sulfide, amide, hydroxyl, amino, and carboxyl groups, the hydrogen-bond-formable compound (HC) contains at least one functional group selected from the group consisting of sulfide, amide, hydroxyl, amino, and carboxyl groups, the at least one functional group in the prot