Container for administering, storing, transporting or transporting protein or protein-containing composition with low protein adsorption, and apparatus for manufacturing protein or protein composition

Abstract

The present application relates to a container for administering, storing, handling or transporting a protein or a protein-containing composition with low protein adsorption and a device for manufacturing a protein or a protein composition. A container in which a surface that comes into contact with a protein or a protein-containing composition is formed from the following fluororesin, and a device for producing a protein or a protein-containing composition have significantly low protein adsorbability, the fluororesin is at least one type of fluororesin selected from the group consisting of tetrafluoroethylene-hexafluoropropylene copolymers and tetrafluoroethylene-perfluoroalkyl vinyl ether copolymers, the melting point of the fluororesin is 320 DEG C or less, and the total number of non-fluorinated group terminals and-CF2H group terminals per 1 * 106 carbon atoms in the fluororesin is 70 or less.

Description

[0001] This application is a divisional application of Chinese Patent Application No. 201780083399.6 (PCT Application No. PCT / JP2017 / 045893) filed on December 21, 2017, entitled "Container for administering, storing, carrying, or transporting a protein or a composition containing a protein with low protein adsorption and apparatus for manufacturing a protein or a protein composition." TECHNICAL FIELD

[0002] The present application relates to a container for administering, storing, carrying, or transporting a protein or a composition containing a protein and an apparatus for manufacturing a protein or a protein composition, characterized in that a surface in contact with a protein or a composition containing a protein is formed of a fluororesin selected from at least one fluororesin among a tetrafluoroethylene-hexafluoropropylene-based copolymer and a tetrafluoroethylene-perfluoroalkyl vinyl ether-based copolymer, and the melting point of the fluororesin is 320°C or lower, and the total number of non-fluorinated group terminals and -CF2H group terminals per 1 x 10 6 carbon atoms in the fluororesin is 70 or lower. BACKGROUND

[0003] In the fields of medical, pharmaceutical, agricultural, biological, and the like research, or the fields of manufacturing technology of protein preparations such as pharmaceutical products, especially antibody pharmaceutical products, and the field of regenerative medicine, a protein itself and a composition containing a protein are frequently handled.

[0004] Proteins play an important role in maintaining life by transmitting information and producing and transporting physiologically active substances. However, when proteins and compositions containing proteins are used in the above research fields and technical fields, problems often arise due to adsorption of the proteins. For example, in the manufacturing (culturing, purification, etc.) process and the storage and transport process of protein preparations such as antibody drugs, the manufacturing cost increases when the protein preparations are lost due to adsorption to equipment. In addition, when protein preparations such as antibody drugs are administered, the actual amount administered will be less than the amount indicated on the container if the protein preparations are lost due to adsorption to equipment, so the therapeutic effect can also be affected. In addition, in the cell culturing process aimed at regenerative medicine, cell research, etc., there is a problem that the cost increases when expensive protein components (growth factors required for cell growth, differentiation induction, etc.) contained in the culture medium (particularly, serum-free culture medium, differentiation-inducing culture medium) are lost due to adsorption to equipment during culturing. Furthermore, irreversible adsorption of proteins becomes a cause of contamination in chromatography columns and experimental tubes, and in the case of hemodialysis membranes, etc., not only does activation of the complement system occur, but the inherent membrane permeability also significantly decreases, and the function of material exchange, etc. cannot be sufficiently exerted. Furthermore, activation of cells, immune responses, etc. are induced, and the material will soon be recognized as a foreign object.

[0005] Recently, in the technical field of pharmaceuticals, the importance of protein preparations such as antibody drugs is increasing. On the other hand, there has also been progress in the practical use of regenerative medicine using various cells, tissues, etc. containing iPS cells, cell sheets. Therefore, in various fields, there is an urgent need for materials that interact weakly with proteins in nature and do not adsorb proteins.

[0006] Prior to this, various materials having low adsorptivity with respect to proteins and medical and experimental instruments (containers, syringes, catheters, laboratory instruments, devices for treatment, etc.) using such materials have been proposed. For example, as a non-fluorinated polymer for the purpose of preventing adsorption of biological related substances such as proteins, an ethylene-vinyl alcohol copolymer (Patent Document 1); a polyurea-urethane polymer (Patent Document 2); a mixture of a water-soluble copolymer and a hydrazide compound having at least two hydrazine groups per molecule (Patent Document 3); a copolymer formed of a plurality of repeating units (Patent Documents 4 and 5); a hydrophilized oil or a hydrophilized copolymer (Patent Document 6); a blend of a water-soluble polymer and a base polymer (Patent Document 7); a copolymer characterized by being a copolymer of 2-methacryloyloxyethylphosphorylcholine, n-butyl (meth) acrylate, methyl (meth) acrylate, or styrene, and being soluble in a buffer and physiological saline (Patent Document 8); a copolymer characterized by being a copolymer containing repeating units derived from an ethylenically unsaturated polymerizable monomer (a) having an alkylene glycol residue and an ethylenically unsaturated polymerizable monomer (b) having a functional group for fixing a physiologically active substance bonded via an alkylene glycol residue, and having a reactive functional group at at least one terminal end of the copolymer (Patent Document 9); a cyclic olefin resin (Patent Document 10); and the like have been proposed.

[0007] In addition, in the case where a fluororesin is used as a material having low adsorptivity with respect to proteins, as a protein adhesion preventing compound capable of forming a coating layer excellent in water resistance, from which a coating component is not easily eluted, and in which proteins are not easily adsorbed, a coating liquid using the same, and a medical device using the same, a protein adhesion preventing compound characterized by being a protein adhesion preventing compound for forming a coating layer for preventing adsorption of proteins on the surface of an article, which contains a fluorine-containing polymer, and a medical device having a coating layer formed of the protein adhesion preventing compound on the surface are proposed (Patent Document 11). In Patent Document 11, it is described that "the fluorine atom content of the fluorine-containing polymer is preferably 5 to 90 mass%, more preferably 10 to 85 mass%, and particularly preferably 15 to 80 mass%. When the fluorine atom content is equal to or higher than the lower limit of the foregoing range, the water resistance is excellent. When the fluorine atom content is equal to or lower than the upper limit of the foregoing range, proteins are not easily adsorbed." (

[0013] paragraph). In Patent Document 11, various fluororesins including FEP, PFA are listed, and as an example, it is described that a coating liquid obtained by dissolving FEP having a fluorine atom content of 76.0% in chloroform is used to form a film layer on the surface of a hole.

[0008] Prior Art Documents

[0009] Patent Documents

[0010] Patent Literature 1: Japanese Patent Application Laid-Open No. 1-213137

[0011] Patent Literature 2: Japanese Patent Application Laid-Open No. 5-103831

[0012] Patent Literature 3: Japanese Patent No. 4941672

[0013] Patent Literature 4: Japanese Patent No. 5003902

[0014] Patent Literature 5: Japanese Patent No. 5207012

[0015] Patent Literature 6: Japanese Patent Application Laid-Open No. 2002-505177

[0016] Patent Literature 7: Japanese Patent Application Laid-Open No. Hei 7-502563

[0017] Patent Literature 8: Japanese Patent No. 3443891

[0018] Patent Literature 9: Japanese Patent Application Laid-Open No. 2008-1794

[0019] Patent Literature 10: Japanese Patent Application Laid-Open No. 2016-155327

[0020] Patent Literature 11: Japanese Patent Application Laid-Open No. 2016-26520 SUMMARY

[0021] PROBLEMS TO BE SOLVED BY THE INVENTION

[0022] For the apparatuses described in Patent Literatures 1 to 10 described above, which are manufactured using non-fluorinated polymer materials, durability and oil resistance are insufficient, and from a practical viewpoint, it cannot be said that the desired effects of low protein adsorption are sufficiently exerted.

[0023] Further, for the apparatus described in Patent Literature 11 described above, which is manufactured using a fluorinated base material, even in the case where FEP is used, there is a problem that low protein adsorption is insufficient.

[0024] The present application has an object to solve the problems of the conventional technologies described above, and to provide a container for application, storage, transportation, or transportation of a protein or a composition containing a protein, and an apparatus for manufacturing a protein or a protein composition, which can sufficiently exert low protein adsorption.

[0025] MEANS FOR SOLVING THE PROBLEMS

[0026] The present inventors have conducted intensive studies in order to solve the above problems, and as a result, have found that, when a container having a surface in contact with a protein formed of a fluororesin selected from at least one fluororesin of a tetrafluoroethylene-hexafluoropropylene copolymer and a tetrafluoroethylene-perfluoroalkyl vinyl ether copolymer and having a melting point of 320°C or lower and in which the total number of non-fluorinated group terminals and -CF2H group terminals per 1 x 10 6 carbon atoms is 70 or less is used to administer, store, carry or transport a protein or a composition containing a protein, or a component for producing a protein composition formed of a fluororesin selected from at least one fluororesin of a tetrafluoroethylene-hexafluoropropylene copolymer and a tetrafluoroethylene-perfluoroalkyl vinyl ether copolymer and having a melting point of 320°C or lower and in which the total number of non-fluorinated group terminals and -CF2H group terminals per 1 x 10

[0027] That is, the present application is as described below.

[0028] (1) A container for administering, storing, carrying or transporting a protein or a composition containing a protein, or a device for producing a protein or a composition containing a protein, characterized in that a surface in contact with a protein or a composition containing a protein is formed of a fluororesin selected from at least one fluororesin of a tetrafluoroethylene-hexafluoropropylene copolymer and a tetrafluoroethylene-perfluoroalkyl vinyl ether copolymer and having a melting point of 320°C or lower and in which the total number of non-fluorinated group terminals and -CF2H group terminals per 1 x 10 6 carbon atoms is 70 or less.

[0029] (2) The container for administering, storing, carrying or transporting a protein or a composition containing a protein, or the device for producing a protein or a composition containing a protein according to (1), characterized in that it is a container.

[0030] (3) The container for administering, storing, carrying or transporting a protein or a composition containing a protein, or the device for producing a protein or a composition containing a protein according to (1) or (2), characterized in that it is a bag.

[0031] (4) The container for administering, storing, carrying or transporting a protein or a composition containing a protein, or the device for producing a protein or a composition containing a protein according to any one of (1) to (3), characterized in that the protein or the composition containing a protein is an antibody (immunoglobulin).

[0032] (5) The container for administering, storing, carrying or transporting a protein or a composition containing a protein or the apparatus for producing a protein or a composition containing a protein according to any one of (1) to (3), characterized in that the protein or the composition containing a protein is albumin.

[0033] (6) The container for administering, storing, carrying or transporting a protein or a composition containing a protein or the apparatus for producing a protein or a composition containing a protein according to (1), characterized in that it is an apparatus for producing a protein preparation.

[0034] Effects of the Invention

[0035] For the fluororesin of the present application, especially FEP·PFA in which the total number of non-fluorinated group terminals and -CF2H group terminals is 70 or less per 1 x 10 6 Since the FEP·PFA in which the total number of non-fluorinated group terminals and -CF2H group terminals is 70 or less per 1 x 10

[0036] (1) In the production (culturing, purification, etc.) process, the storage and transport process or the administration of a protein preparation such as an antibody drug, loss due to adsorption of the protein preparation to the apparatus is prevented.

[0037] (2) In the cell culturing process (including the differentiation induction process) in regenerative medical use, etc., loss due to adsorption of expensive protein components (growth factors, etc. required for growth, differentiation induction, etc. of cells, specifically, various proteins (albumin, insulin, transferrin, etc. cell growth factors, activin A, bone morphogenetic factor 4 (BMP-4), epithelial growth factor (EGF), stem cell factor (SCF), interleukins, etc. cytokines, growth factors, etc.) contained in the culture medium (especially serum-free culture medium, differentiation induction culture medium) to the apparatus in the culturing is prevented (cost reduction is achieved). DETAILED DESCRIPTION

[0038] As the container for administration, storage, transport or conveyance of a protein or a composition containing a protein, or the apparatus for manufacturing a protein or a composition containing a protein (hereinafter, sometimes simply referred to as "the container or apparatus of the present application" or "the container or apparatus") of the present application, there is no particular limitation as long as it is a container or apparatus for administration, storage, transport or conveyance of a protein or a composition containing a protein, or for manufacturing a protein or a composition containing a protein, which has a surface in contact with a protein or a composition containing a protein formed of a fluororesin (hereinafter, these fluororesins will be collectively referred to as "the fluororesin of the present application") selected from at least one fluororesin among tetrafluoroethylene-hexafluoropropylene copolymer and tetrafluoroethylene-perfluoroalkyl vinyl ether copolymer, and the melting point of the fluororesin is 320°C or lower, and the total number of non-fluorinated group terminals and -CF2H group terminals in the fluororesin is 70 or lower per 1 x 10 6 carbon atom. The container or apparatus of the present application is characterized in that the surface of the container or apparatus in contact with a protein or a composition containing a protein is formed of the fluororesin of the present application. When administration, storage, transport or conveyance of a protein (e.g., an antibody (immunoglobulin)) or a composition containing a protein is performed using the container or apparatus having the above-described characteristics, for example, in the manufacturing (culturing, purification, etc.) process, the storage and transport process or the administration of a protein preparation such as an antibody drug, loss due to adsorption of the protein preparation to the apparatus, loss due to adsorption of an expensive protein component (e.g., a growth factor, a cytokine, etc. required for growth, differentiation induction, etc. of cells) to the apparatus can be prevented, and thus cost reduction is achieved.

[0039] In the present application, the "protein" means one or more kinds of high molecular compounds in which a plurality of L-amino acids are linked (polymerized) by amide bonds (also referred to as peptide bonds) in a chain shape, and the number of amino acids as a constituent element is not limited. Therefore, the so-called peptide is also included in the protein of the present application. In addition, glycoprotein in which a sugar is bonded to a protein, and lipoprotein in which a lipid is bonded to a protein are also included in the protein of the present application. As the protein used in the present application, albumin, fibrinogen, globulin (al-globulin, a2-globulin, β-globulin, γ-globulin), erythropoietin, collagen, elastin, keratin, lactoferrin, avidin, cadherin, proteoglycan, mucin, LDL (Low Density Lipoprotein), HDL (High Density Lipoprotein), VLDL (Very Low Density Lipoprotein), insulin, a cell growth factor such as transferrin, Activin A, Bone Morphogenetic Protein 4 (BMP-4), Epidermal Growth Factor (EGF), Stem Cell Factor (SCF), a cytokine such as interleukin, and a growth factor can be exemplified, but are not limited to these.

[0040] In the present application, the "protein-containing composition" means a mixture or a product of one or more kinds of proteins and one or more kinds of other substances. As the protein-containing composition used in the present application, a protein preparation such as an antibody drug, a body fluid such as blood, a biological component containing a protein such as serum, plasma, a culture medium containing a protein component (particularly, a serum-free culture medium, a differentiation-inducing culture medium), and the like can be exemplified, but are not limited to these.

[0041] In the present application, the "container for administration", the "container for storage", the "container for transport", the "container for transport", the "apparatus for manufacturing", and the "apparatus" in the container for administering, storing, transporting, or transporting a protein or a protein-containing composition each have the following meanings.

[0042] The "container for administration" means a container used when a protein or a protein-containing composition is administered to a patient in a clinic.

[0043] The "container for storage" means a container used when a protein or a protein-containing composition is stored for a certain period of time.

[0044] The "container for carrying" means a container used when a protein or a composition containing a protein is moved by a human power or a machine (including a robot) or the like.

[0045] The "container for transportation" means a container used when a protein or a composition containing a protein is transferred by a transportation means such as a car, a ship, an airplane or the like.

[0046] The "equipment for manufacturing" means an equipment used when a protein or a composition containing a protein is manufactured.

[0047] The "equipment" means an implement (a simple tool), an instrument (a device, a tool which is directly manipulated by a human, is relatively small, and is small-scale), and a material for manufacturing the implement and the instrument. For example, a pipe, a tube, a container or the like of a manufacturing apparatus for an antibody drug or the like, and an equipment for purification (a filter, a column or the like) can be exemplified.

[0048] For the fluororesin of the present application, the total of the non-fluorinated group terminal (for example, -COF, -COOH, and -COOH associated with water, -CH2OH, -CONH2, -COOCH3, and the like) and the -CF2H group terminal in the fluororesin is preferably 70 or less, more preferably 35 or less, further more preferably 20 or less, particularly preferably 10 or less, per 1 x 10 6 carbon atoms. The fluororesin can be one which does not have the -CF2H group terminal. In the case where the fluororesin does not have the -CF2H group terminal, the total of the non-fluorinated group terminal in the fluororesin is preferably 70 or less, more preferably 35 or less, further more preferably 20 or less, particularly preferably 10 or less, per 1 x 10 6 carbon atoms. The fluororesin can be one which does not have the -CF2H group terminal. In the case where the fluororesin does not have the -CF2H group terminal, the total of the non-fluorinated group terminal in the fluororesin is preferably 70 or less, more preferably 35 or less, further more preferably 20 or less, particularly preferably 10 or less, per 1 x 10 6 carbon atoms. The fluororesin can be one which does not have the -CF2H group terminal. In the case where the fluororesin does not have the -CF2H group terminal, the total of the non-fluorinated group terminal in the fluororesin is preferably 70 or less, more preferably 35 or less, further more preferably 20 or less, particularly preferably 10 or less, per 1 x 10 6 carbon atoms. The fluororesin can be one which does not have the -CF2H group terminal. In the case where the fluororesin does not have the -CF2H group terminal, the total of the non-fluorinated group terminal in the fluororesin is preferably 70 or less, more preferably 35 or less, further more preferably 20 or less, particularly preferably 10 or less, per 1 x 10 6 carbon atoms. The fluororesin can be one which does not have the -CF2H group terminal. In the case where the fluororesin does not have the -CF2H group terminal, the total of the non-fluorinated group terminal in the fluororesin is preferably 70 or less, more preferably 35 or less, further more preferably 20 or less, particularly preferably 10 or less, per 1 x 10 6 carbon atoms. The fluororesin can be one which does not have the -CF2H group terminal. In the case where the fluororesin does not have the -CF2H group terminal, the total of the non-fluorinated group terminal in the fluororesin is preferably 70 or less, more preferably 35 or less, further more preferably 20 or less, particularly preferably 10 or less, per 1 x 10 6 carbon atoms. The fluororesin can be one which does not have the -CF2H group terminal. In the case where the fluororesin does not have the -CF2H group terminal, the total of the non-fluorinated group terminal in the fluororesin is preferably 70 or less, more preferably 35 or less, further more preferably 20 or less, particularly preferably 10 or less, per 1 x 10 6 carbon atoms. The fluororesin can be one which does not have the -CF2H group terminal. In the case where the fluororesin does not have the -CF2H group terminal, the total of the non-fluorinated group terminal in the fluororesin is preferably 70 or less, more preferably 35 or less, further more preferably 20 or less, particularly preferably 10 or less, per 1 x 10

[0049] Note that the number of the above -COF, -COOH, and -COOH associated with water, -CH2OH, -CONH2, -COOCH3, and -CF2H per 1 x 10 6 carbon atoms can be calculated from FT-IR.

[0050] In the present application, the "non-fluorinated group terminal" refers to a terminal having reactivity, which is generally called a labile terminal, and as the non-fluorinated group terminal, specifically, functional groups such as -COF, -COOH, -COOH associated with water, -CH2OH, -CONH2, -COOCH3, and the like can be cited.

[0051] The melting point of the fluororesin of the present application is 320°C or lower, and 240°C or higher. As a preferable melting point range, for example, 245°C or higher and 315°C or lower, 250°C or higher and 310°C or lower can be cited.

[0052] As the fluororesin of the present application, specifically, tetrafluoroethylene (TFE)-hexafluoropropylene (HFP) copolymer (FEP), TFE-perfluoroalkyl vinyl ether (PAVE) copolymer (PFA) can be cited.

[0053] In the above, the FEP·PFA in which the total number of the non-fluorinated group terminal and the -CF2H group terminal per 1 x 10 6 The FEP·PFA in which the total number of the non-fluorinated group terminal and the -CF2H group terminal per 1 x 10 6 The FEP·PFA in which the total number of the non-fluorinated group terminal and the -CF2H group terminal per 1 x 10 6 The FEP·PFA in which the total number of the non-fluorinated group terminal and the -CF2H group terminal per 1 x 10 6 The FEP·PFA in which the total number of the non-fluorinated group terminal and the -CF2H group terminal per 1 x 10 6 The FEP·PFA in which the total number of the non-fluorinated group terminal and the -CF2H group terminal per 1 x 10 6 The FEP·PFA in which the total number of the non-fluorinated group terminal and the -CF2H group terminal per 1 x 10

[0054] The fluororesin of the present application has the following (1) to (5) in addition to the above-described characteristics.

[0055] (1) No elution of plasticizers or the like.

[0056] (2) High-temperature steam (autoclave) sterilization can be performed.

[0057] (3) Insoluble in DMSO, DMF.

[0058] (4) Excellent super low temperature characteristics (not brittle even at -200°C).

[0059] (5) High transparency.

[0060] The "TFE-HFP copolymer" described above refers to a copolymer containing at least TFE and HFP. That is, in the "TFE-HFP copolymer", in addition to a binary copolymer of TFE and HFP (TFE / HFP copolymer; FEP), a ternary copolymer of TFE and HFP and vinyl fluoride (VF) (TFE / HFP / VF copolymer), TFE and HFP and vinylidene fluoride (VDF) (TFE / HFP / VDF copolymer), TFE and HFP and perfluoro(alkyl vinyl ether) (PAVE) (TFE / HFP / PAVE copolymer), and the like, a quaternary copolymer of TFE and HFP and VF and VDF (TFE / HFP / VF / VDF copolymer), TFE and HFP and VF and PAVE (TFE / HFP / VF / PAVE copolymer), TFE and HFP and VDF and PAVE (TFE / HFP / VDF / PAVE copolymer), and the like, a quinary copolymer of TFE and HFP and VF and VDF and PAVE (TFE / HFP / VF / VDF / PAVE copolymer), and the like are also included.

[0061] In the fluororesin of the present application, the melting point of FEP is 300°C or lower, and 240°C or higher. As a preferable melting point range, for example, 245°C or higher and 290°C or lower, 250°C or higher and 280°C or lower can be given.

[0062] As the TFE-HFP copolymer described above, a TFE / HFP copolymer, a TFE / HFP / PAVE copolymer are preferable. The mass ratio of TFE to HFP in the TFE / HFP copolymer is preferably 80 to 97 / 3 to 20, more preferably 84 to 92 / 8 to 16. Further, the mass ratio of TFE to HFP to PAVE in the TFE / HFP / PAVE copolymer is preferably 70 to 97 / 3 to 20 / 0.1 to 10, more preferably 81 to 92 / 5 to 16 / 0.3 to 5.

[0063] The "TFE-PAVE-based copolymer" described above refers to a copolymer containing at least TFE and PAVE. That is, in the "TFE-PAVE-based copolymer", in addition to a binary copolymer of TFE and PAVE (TFE / PAVE copolymer; PFA), there are included a copolymer of TFE and PAVE and hexafluoropropylene (HFP) (TFE / PAVE / HFP copolymer), a copolymer of TFE and PAVE and vinylidene fluoride (VDF) (TFE / PAVE / VDF copolymer), a copolymer of TFE and PAVE and chlorotrifluoroethylene (CTFE) (TFE / PAVE / CTFE copolymer), and the like, a ternary copolymer, a copolymer of TFE and PAVE and HFP and VDF (TFE / PAVE / HFP / VDF copolymer), a copolymer of TFE and PAVE and HFP and CTFE (TFE / PAVE / HFP / CTFE copolymer), a copolymer of TFE and PAVE and VDF and CTFE (TFE / PAVE / VDF / CTFE copolymer), and the like, a quaternary copolymer, and a copolymer of TFE and PAVE and HFP and VDF and CTFE (TFE / PAVE / HFP / VDF / CTFE copolymer) and the like, a quinary copolymer.

[0064] As the PAVE constituting the PAVE unit described above, there is no particular limitation, and examples that can be given include perfluoro(methyl vinyl ether) [PMVE], perfluoro(ethyl vinyl ether) [PEVE], perfluoro(propyl vinyl ether) [PPVE], perfluoro(butyl vinyl ether), perfluoro(pentyl vinyl ether), perfluoro(hexyl vinyl ether), perfluoro(heptyl vinyl ether), and the like.

[0065] In the fluororesin of the present application, the melting point of PFA is 320°C or lower and 285°C or higher. As a preferable range of the melting point, for example, 290°C or higher and 315°C or lower, 295°C or higher and 315°C or lower, and 300°C or higher and 310°C or lower can be given.

[0066] The mass ratio of TFE to PAVE in the TFE-PAVE-based copolymer described above is preferably 90 to 98 / 2 to 10, and more preferably 92 to 97 / 3 to 8.

[0067] The fluororesin of the present application can be produced by fluorination treatment of the terminal groups of a fluororesin synthesized according to a conventional method such as suspension polymerization or emulsion polymerization, using a method known per se such as a method in which the fluororesin is stabilized by contact with a fluorine-containing compound (for example, a fluorine radical source) before melt extrusion of the fluororesin, a method in which the particles of the fluororesin obtained after melt extrusion of the fluororesin are fluorinated by contact with a fluorine-containing compound, and the like. In addition, elemental fluorine, a chain transfer agent capable of controlling the terminal groups, and a polymerization catalyst can be used in the production of the fluororesin (at the time of polymerization). In addition, commercially available products can be used as the fluororesin of the present application. Furthermore, a molded article such as a film obtained by melting and molding the fluororesin, a container or a device molded from the film, a container or a device molded from the fluororesin, and the like can be fluorinated by contact with a fluorine-containing compound. In addition, these treatment methods can be combined.

[0068] That is, the total of the aforementioned non-fluorinated group terminals, and the total of the non-fluorinated group terminals and the -CF2H group terminals need not be 70 or less per 1 x 10 6 carbon atoms at each stage of the fluororesin, the particles, and the film as a raw material, but can be 70 or less per 1 x 10 6 carbon atoms in the surface of the final container or device that comes into contact with a protein. In the case of a fluororesin having one or more -CF3 terminal groups, the -CF3 terminal groups need not be one or more at each stage of the fluororesin, the particles, and the film as a raw material, but the fluororesin can have one or more -CF3 terminal groups in the surface of the final container or device that comes into contact with a protein.

[0069] As the aforementioned fluorine radical source, there is no particular limitation, and fluorinated halogens such as IF5, CIF3, and the like, F2 gas, CoF3, AgF2, UF6, OF2, N2F2, CF3OF, and the like can be mentioned. The F2 gas can be F2 at a concentration of 100%, but from the viewpoint of safety, it is used by being diluted to 5 to 50% by mass, preferably 15 to 30% by mass, by mixing with an inactive gas. As the inactive gas, nitrogen, helium, argon, and the like can be mentioned, and from the viewpoint of cost effectiveness, nitrogen is preferred.

[0070] The aforementioned fluorination treatment is preferably performed at a temperature of 20 to 220°C, more preferably at a temperature of 100 to 200°C. The aforementioned fluorination treatment is preferably performed for 5 to 30 hours, more preferably for 10 to 20 hours.

[0071] The container or apparatus obtained by the present application can be one in which the arithmetic average roughness (Ra) of the surface roughness, the root mean square roughness (RMS) of the surface roughness, and the surface free energy have been adjusted. For example, a container or apparatus in which the Ra of the surface roughness is 3.5 to 6.5 nm, the RMS of the surface roughness is 4.5 to 8.0 nm, and the surface free energy is 16.5 to 18.5 (mJ / m 2 ) can be mentioned.

[0072] As described above, the FEP·PFA in which the total number of non-fluorinated group terminals and -CF2H group terminals is 70 or less per 1 x 10 6 carbon atoms has extremely superior properties compared to the FEP·PFA in which the total number of non-fluorinated group terminals and -CF2H group terminals is more than 70 per 1 x 10 6 carbon atoms, and in particular has the following advantages.

[0073] (1) In the manufacturing (culturing, purification, etc.) process, the storage and transport process, or at the time of administration of a protein preparation such as an antibody drug, loss due to adsorption of the protein preparation to the apparatus is prevented.

[0074] (2) In the cell culturing process (including the differentiation induction process) in regenerative medical applications, etc., loss (leading to cost reduction) due to adsorption of expensive protein components (growth factors, cytokines, etc. required for cell growth, differentiation induction, etc.) contained in the culture medium (particularly serum-free culture medium, differentiation induction medium) to the apparatus during culturing is prevented.

[0075] The fluororesin of the present application can be used for various containers, parts of manufacturing equipment, apparatuses for purification, experimental apparatuses, etc. having a surface that comes into contact with a protein or a composition containing a protein.

[0076] As the form of the container or apparatus of the present application, for example, a bag, a bottle, a centrifuge tube, a vial, a syringe, a tube, etc. can be mentioned. In the case where the container of the present application is a container for administration of a protein, a syringe, a (drip) bag, a (drip) bottle, a tube are preferred, in the case where the container of the present application is a container for storage of a protein, a bag, a bottle, a centrifuge tube, a vial are preferred, and in the case where the container of the present application is a container for transport and transport of a protein, a bag, a bottle, a vial, a tube are preferred. In particular, the container of the present application in the form of a bag can be applied to all of the uses for administration, storage, transport, and transport of a protein, and thus can be preferably exemplified. As the specific use of the manufacturing apparatus of the present application, for example, the following uses can be mentioned.

[0077] (1) Protein preparation related to an antibody drug, etc.:

[0078] Culture containers (bags, etc.), piping of manufacturing equipment, containers for use in applications or storage, purification equipment (filters, columns, etc.), containers for preservation / transport, containers for administration (syringes, administration bags, etc.)

[0079] (2) Cell culture-related containing protein components in regenerative medical applications and the like:

[0080] Culture containers (bags, etc.) (particularly for mass culture of iPS cells, for differentiation induction), culture medium containers (including containers for protein components such as proliferation / growth factors, cytokines, etc.)

[0081] The above bags, bottles, centrifuge tubes, vials, syringes, tubes, etc. can be manufactured by combining molding methods such as compression molding, extrusion molding, transfer molding, blow molding, blow molding, injection molding, rotational molding, lining molding, foam extrusion molding, film molding, and sealing means such as heat sealing, high-frequency fusion bonding, ultrasonic fusion bonding, etc. as needed. In the case of manufacturing using these methods, there is an advantage that coating work is not required compared to the case of using a coating agent for coating.

[0082] The above bag can be manufactured by, for example, overlapping the film (sheet) of the fluororesin raw material of the present application and heat-sealing the edge portion using a pulse heat sealer.

[0083] The film used in the molding of the above bag can be a single-layer film or a laminated film formed of two or more layers. In the case of a laminated film formed of multiple layers, the bag can be molded in such a manner that the inner surface that comes into contact with the mammalian cells is a layer film of the fluororesin raw material of the present application, and the other layer films can be layer films of raw materials (for example, polyolefin-based resin raw materials) other than the fluororesin of the present application. The lamination of the film is performed using methods such as heat lamination, heat compression, high-frequency heating, solvent casting, and extrusion lamination.

[0084] In addition, a container or equipment of the present application can also be obtained by performing a coating treatment on a substrate such as a bag, a bottle, a centrifuge tube, a vial, a syringe, a tube, etc. manufactured of glass, metal, resin, etc. using a coating agent formed of the fluororesin of the present application. Any method can be used depending on the form of the substrate. As the coating treatment, methods such as spin coating, spray coating, bar coating, roll coating, immersion, brush coating, spin lining, electrostatic coating, etc. can be mentioned. After coating the fluororesin coating agent described above on the substrate, a coating layer can be formed using drying treatment and high-temperature heating treatment. In addition, the coating layer can be thickened to an arbitrary film thickness by further applying a coating agent containing the fluororesin of the present application.

[0085] Hereinafter, the present application will be further specifically described by way of examples, but the technical scope of the present application is not limited by these examples.

[0086] Example 1

[0087] 1. Container manufacturing

[0088] For five types of films with dimensions of 10cm×4cm and a thickness of 100μm, two films were overlapped and heat-sealed using a pulse heat sealer under the conditions of a sealing time of 50 seconds, a sealing pressure of 0.2MPa, and a sealing width of 4mm, thereby manufacturing five types of perfluoropolymer bags (containers A to E).

[0089] It should be noted that, as a polyethylene bag (container F), a commercially available bag with dimensions of 70×50×0.04mm (unipac (registered trademark) A-4 manufactured by Japan Co., Ltd.) is used, and as a glass container (container G), a commercially available threaded tube bottle with a tube diameter of φ21mm and a total length of 45mm (9mL TS threaded tube bottle manufactured by Maruemu Corporation) is used.

[0090] 2. Determination of the number of non-fluorinated group terminals and the number of -CF2H terminals

[0091] Samples of the resin with a thickness of approximately 250–300 μm were prepared and analyzed using an FT-IR spectrometer 1760X (manufactured by Perkin-Elmer).

[0092] When preparing samples of this resin with a thickness of approximately 250–300 μm, the film constituting the bag (made from granules by melt molding) is measured directly. If the thickness is insufficient, the films are overlapped and measured.

[0093] Obtain the differential spectrum with the standard sample (a sample that has been fully fluorinated until no substantial difference can be observed in the spectrum), read the absorbance of each peak, and calculate the absorbance relative to each 1×10⁻⁶ peak using the following formula. 6 The number of non-fluorinated group terminals and the number of CF2H terminals per carbon atom. The number of non-fluorinated group terminals and the number of CF2H terminals for each bag are shown in Table 2.

[0094] Number of non-fluorinated group ends and -CF2H ends (per 1×10) 6 (number of carbon atoms) = l·k / t

[0095] l: Absorbance

[0096] k: Correction coefficient (see Table 1)

[0097] t: Sample thickness (mm)

[0098] [Table 1]

[0099] Table 1: Absorption wavenumbers and correction factors for each non-fluorinated terminal group and -CF2H terminal group

[0100]

[0101] [Table 2]

[0102] Table 2: Non-fluorinated group end number and -CF2H end number of containers

[0103] Container name Material of container Number of non-fluorinated group ends - CF2H end number A FEP 21 424 B FEP 13 0 C FEP 68 0 D PFA 201 159 E PFA 25 0 F Polyethylene - - G Glass - -

[0104] Example 2

[0105] (Protein non-adhesion)

[0106] (1) Preparation of color developing solution, protein solution

[0107] As the color developing solution, a product obtained by mixing 50 mL of peroxidase color developing solution (3, 3', 5, 5'-tetramethylbenzidine (TMBZ), manufactured by KPL) and 50 mL of TMB peroxidase substrate (manufactured by KPL) was used.

[0108] As the protein solution, a product obtained by diluting protein (POD-goat anti mouse IgG, manufactured by Biorad) to 16,000 times with a phosphate buffer solution (D-PBS, manufactured by Wako Pure Chemical Industries, Ltd.) was used.

[0109] (2) Protein adsorption

[0110] Using a micropipette, 2 mL of the protein solution was injected into each of the containers A to G (2 mL was used for each container), and left at room temperature for 1 hour. Each reaction was performed with N = 3.

[0111] (3) Container washing

[0112] Next, the protein solution was removed from each container, and each container was washed 4 times with 4 mL of a phosphate buffer solution containing 0.05 mass% of a surfactant (TWEEN 20, manufactured by Wako Pure Chemical Industries, Ltd.) (4 mL was used for each container in 4 portions).

[0113] (4) Color developing solution injection

[0114] Next, 2 mL of the color developing solution was injected into each of the containers that had been washed (2 mL was used for each container), and a color developing reaction was performed for 7 minutes. The color developing reaction was terminated by adding 1 mL of a 1M phosphoric acid solution (1 mL was used for each container).

[0115] For the blank, 2 mL of the coloring solution was injected into each of three glass containers (2 mL for each glass container), and then 40 μL of the protein solution was injected into each of the three glass containers. The coloring reaction was performed for 7 minutes, and then the coloring reaction was terminated by adding 1 mL of a 1 M phosphoric acid solution (1 mL for each glass container).

[0116] (5) Absorbance measurement preparation

[0117] Next, 3 mL of the solution was taken from each of the containers and moved to a cuvette for the spectrophotometer.

[0118] (6) Absorbance measurement and protein adsorption rate Q

[0119] For the absorbance, the absorbance at 450 nm was measured using an ultraviolet spectrophotometer U-3310 (manufactured by Hitachi, Ltd.). Here, the average of the absorbances of the blank (N = 3) was denoted as A0. The absorbance of the solution moved from each bag was denoted as A1.

[0120] The protein adsorption rate Q1 was calculated using the following equation, and the average of the protein adsorption rates Q was used.

[0121] Q1 = A1 / {A0 x (2000 / amount of the protein solution injected into the blank)} x 100

[0122] = A1 / {A0 x (2000 / 40)} x 100 [%]

[0123] [Table 3]

[0124]

[0125] As shown in Table 3, with respect to the containers B, C, and E using a perfluoropolymer in which the total number of non-fluorinated group terminals and -CF2H group terminals is 70 or less per 1 x 10 6 carbons, not only compared with the container F of polyethylene and the container G of glass, but also compared with the containers A and D using a perfluoropolymer in which the total number of non-fluorinated group terminals and -CF2H group terminals is more than 70 per 1 x 10 6 carbons, the protein was not easily significantly adsorbed to the surface. That is, the perfluoropolymer in which the total number of non-fluorinated group terminals and -CF2H group terminals is more than 70 per 1 x 10 6 carbons showed a low protein adsorption property of about 1 / 7 to about 1 / 2 of the perfluoropolymer in which the total number of non-fluorinated group terminals and -CF2H group terminals is more than 70 per 1 x 10 6 carbons.

[0126] Example 3

[0127] 1. Manufacture of container H

[0128] As the container H, a container in which a frame line of 10 x 10 mm was drawn on the surface of a cover glass (Matsunami Glass Industries, C025251, 25 x 25 x No. 1) with "Super PAP Pen Liquid Blocker" (Daido Industry) was used.

[0129] 2. Preparation of fluorescently labeled BSA solution

[0130] Using commercially available BSA (Bovine Serum Albumin, Sigma, A7638) and a fluorescent labeling kit (Alexa Fluor(R) 555 NHS Ester, A20009) manufactured by Thermo Fisher, fluorescently (Alexa Fluor(R) 555) labeled BSA was prepared in accordance with the protocol attached to the kit, and the product (fluorescently labeled BSA solution) adjusted to 10 pg / mL was used in the following experiments.

[0131] 3. Adsorption of fluorescently labeled BSA

[0132] Using a micropipette, 1 mL of the fluorescently labeled BSA solution (10 pg / mL) was injected into each of the containers A and B, and left at 37°C for 1 hour. At this time, the liquid contact area of the bag was about 600 mm 2 .

[0133] For the container H, the fluorescently labeled BSA solution was added dropwise with a micropipette so as to be 167 pL / cm 2 inside the frame line made with "Super PAP Pen Liquid Blocker", and was placed in a culture dish, and then left at 37°C for 1 hour.

[0134] Each reaction was performed with N = 3.

[0135] 4. Washing

[0136] After 1 hour, the fluorescently labeled BSA solution was removed from the containers A, B, and H, and then each container was washed with 2 mL of a PBS solution for 4 times.

[0137] 5. Measurement of fluorescence intensity

[0138] For containers A and B, a portion (a square of about 10 x 10 mm) of the portion in contact with the fluorescently labeled protein solution was cut out with scissors, ProLong(R) Diamond Antifade Mountant (manufactured by Thermo Fisher) was dropped thereon, a cover glass was then placed thereon, and observation was performed with a fluorescent microscope (Zeiss, LSM700, x20).

[0139] For container H as well, after washing, ProLong(R) Diamond Antifade Mountant (manufactured by Thermo Fisher) was dropped thereon, a new cover glass was placed thereon, and observation was performed with a fluorescent microscope (Zeiss, LSM700, x20).

[0140] The main observation conditions were as follows:

[0141] • Objective lens: Plan-Apochromat 20X / 0.8 M27

[0142] • Pinhole: 147 μm

[0143] • Number of pixels: 1024 x 1024

[0144] • Laser power: 0.5%

[0145] After observation, analysis was performed with Fiji software, whereby the average fluorescent intensity of the fluorescently labeled BSA adsorbed for each sample was calculated. (For each sample, 5 fields were analyzed respectively)

[0146] The ratio of the average fluorescent intensity of container A and B to that of container H when the average fluorescent intensity of container H was taken as 100 (BSA relative adsorption rate (%)) is shown in Table 4.

[0147] [Table 4]

[0148] Container name Relative adsorption rate of BSA (%) Example 4 B 1.45 Comparative Example 5 A 54.03 Comparative Example 6 H 100

[0149] As shown in Table 4, it was found that, with regard to container B in which a perfluoropolymer in which the total number of non-fluorinated group terminals and -CF2H group terminals was 70 or less per 1 x 10 6 carbon atoms was used, not only was BSA less likely to be significantly adsorbed to the surface compared to container H of glass, but also was BSA less likely to be significantly adsorbed to the surface compared to container A in which a perfluoropolymer in which the total number of non-fluorinated group terminals and -CF2H group terminals was more than 70 per 1 x 10 6 carbon atoms was used.

[0150] Industrial applicability

[0151] The fluororesin of the present application exhibits extremely excellent low protein adsorption, and thus can be used for all machines using a protein or a composition containing a protein. In particular, it can be applied to various devices related to a protein preparation such as an antibody drug, for example, a culture container (bag, etc.), a piping of a manufacturing apparatus, etc., a device for purification (filter · column, etc.), a container for storage / transport, a container for administration (syringe, administration bag, etc.), and various manufacturing devices related to cell culture containing a protein component in a regenerative medical use, etc., for example, a culture container (bag, etc.) (particularly, for mass culture of iPS cells, for differentiation induction), a medium container (including a container for a protein component such as a growth factor), etc.

Claims

1. A container for administering, storing, handling, or transporting proteins or protein-containing compositions, or apparatus for manufacturing proteins or protein-containing compositions, characterized in that: A surface in contact with a protein or a protein-containing composition is formed by a fluororesin, wherein the fluororesin is at least one selected from tetrafluoroethylene-hexafluoropropylene copolymers and tetrafluoroethylene-perfluoroalkyl vinyl ether copolymers, and wherein the melting point of the fluororesin is below 320°C, and the fluororesin contains a certain amount of protein relative to 1 × 10⁻⁶ ppm. 6 The total number of non-fluorinated group ends and -CF2H group ends per carbon atom is less than 70.

2. The container for administering, storing, handling, or transporting proteins or protein-containing compositions as claimed in claim 1, or the apparatus for manufacturing proteins or protein-containing compositions, characterized in that, It is a container.

3. The container for administering, storing, handling, or transporting proteins or protein-containing compositions as described in claim 1 or 2, or the apparatus for manufacturing proteins or protein-containing compositions, characterized in that... It is a bag.

4. The container for administering, storing, handling, or transporting proteins or protein-containing compositions as described in any one of claims 1 to 3, or the apparatus for manufacturing proteins or protein-containing compositions, characterized in that, Proteins or compositions containing proteins are antibodies (immunoglobulins).

5. The container for administering, storing, handling, or transporting proteins or protein-containing compositions as described in any one of claims 1 to 3, or the apparatus for manufacturing proteins or protein-containing compositions, characterized in that, The protein or composition containing the protein is albumin.

6. The container for administering, storing, handling, or transporting proteins or protein-containing compositions as claimed in claim 1, or the apparatus for manufacturing proteins or protein-containing compositions, characterized in that, It is a device used to manufacture protein preparations.

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