JP2010047527A - Method of separating immunoglobulin monomer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method of separating an immunoglobulin monomer with high precision by subjecting an immunoglobulin solution containing an immunoglobulin monomer, an aggregate thereof, and a protein having an isoelectric point lower than the isoelectric point of the immunoglobulin to crossflow filtration with the use of an ultrafiltration membrane, a module of the ultrafiltration membrane, and a crossflow filter. <P>SOLUTION: The method of separating an immunoglobulin monomer includes subjecting an immunoglobulin solution containing at least an immunoglobulin monomer, an aggregate thereof, and a protein having an isoelectric point of lower than the isoelectric point of the immunoglobulin and having a concentration of the immunoglobulin of 1-150 g/L and a pH of not lower than the isoelectric point of the immunoglobulin to crossflow filtration with the use of an ultrafiltration membrane having a cut-off molecular weight of 100,000 to less than 500,000. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention uses an ultrafiltration membrane to cross-flow filter an immunoglobulin solution containing at least an immunoglobulin monomer, an aggregate thereof, and a protein having an isoelectric point lower than that of the immunoglobulin. The present invention relates to a method of separating the water with high accuracy, an ultrafiltration membrane module thereof, and a crossflow filtration device.

Immunoglobulins (antibodies) are mainly present in blood and body fluids, and recognize and bind to microorganisms such as bacteria and viruses that have entered the body and cells infected with microorganisms as antigens. When an antibody binds to an antigen, the phagocytic cells such as leukocytes and macrophages recognize and phagocytose the antigen-antibody complex and work to remove it from the body, or immune cells such as lymphocytes bind to cause an immune reaction. Or Thus, immunoglobulins play an important role in infection defense mechanisms.

As a method for producing an immunoglobulin, there are a method of separating and purifying from biological components, mainly blood, and a method of separating and purifying from cells such as hybridomas. However, there are 100,000 or more kinds of microorganisms such as proteins, DNA, RNA, viruses and the like as biological components, and it is necessary to separate them from immunoglobulins. A protein having a particularly high content is albumin, and separation and purification of immunoglobulin and albumin is an important issue.
In addition, there are aggregates (mainly dimers) in which a plurality of immunoglobulins are bound together by non-covalent bonds. Immunoglobulin aggregates are considered to be one of the causes of side effects that appear when an immunoglobulin preparation is intravenously injected into the human body, such as shock-like reactions such as cyanosis and decreased blood pressure, respiratory tract symptoms such as dyspnea, and rashes. Yes. These aggregates tend to aggregate with each other to form larger multimers, which can cause cloudiness and precipitation in the solution. In some cases, other proteins, bacteria, viruses, and the like are used as nuclei and bound around them to form huge protein aggregates called antigen-antibody complexes. Once formed, aggregates generally cannot be easily dissociated. Therefore, removal of these aggregates from the solution is cited as an important issue.

Several purification methods have been reported to separate immunoglobulin monomers from the solution in which they are mixed. Examples include ion exchange chromatography, hydrophobic chromatography, gel chromatography, chemical treatment, adsorption, and ultrafiltration membrane.
Ion exchange chromatography and hydrophobic chromatography methods are effective when separating substances with greatly different charges and hydrophobicity, but substances with similar degrees of charge and hydrophobicity, such as immunoglobulin monomers and dimers. However, there are disadvantages such as not being able to separate sufficiently and requiring a large amount of eluent and salt.
On the other hand, the gel chromatography method is effective in separating immunoglobulin dimers and dimers having different sizes because of size separation, but cannot process a large amount of immunoglobulins and requires work. There is a disadvantage that the time and cost are long.

Chemical treatment methods that add chemical substances can be processed in large quantities, but it is necessary to remove the chemicals used in the treatment from the solution. This will reduce the transmittance.
Also, the adsorption method does not have high removal efficiency of the immunoglobulin dimer, and further, the work of removing the added adsorbent is required in the same manner as the chemical treatment.

  Recently, an ultrafiltration membrane method that has high robustness, is simple, and can process a large amount of protein has attracted attention, and there are many reports. In general, when separating multiple components by ultrafiltration, the fractionation performance and permeation amount are considered to be affected by the pore size and charge state in the cake layer formed on the surface of the ultrafiltration membrane. For the formation of the cake layer, the fractional molecular weight of the ultrafiltration membrane and the concentration of particles such as proteins constituting the cake layer are presumed as important factors. However, since analysis of the cake layer is difficult, little is known about how to control the cake layer.

  As an immunoglobulin membrane separation method, a globulin dimer removal method (Patent Document 1) by filtering with an ultrafiltration membrane formed from a polysulfone polymer has been proposed. However, since this method is a dead-end method, a large amount of immunoglobulin is trapped in the membrane and a thick cake layer is formed, resulting in membrane clogging. As a result, the transmittance of the immunoglobulin monomer is as low as about 40%, the filtration rate and the filtration capacity are low, and it cannot be said that the industrial effectiveness is high.

  In addition, a method for removing immunoglobulin aggregates by dead-end filtration using a regenerated cellulose hollow fiber membrane (Patent Document 2) has also been proposed, but the immunoglobulin dimer has a large pore diameter (fractional molecular weight). Was hardly removed, and it was not sufficient as a method for separating immunoglobulin dimers.

  When there are a lot of impurities in the solution, the cross-flow filtration method in which membrane clogging hardly occurs is often used in consideration of membrane trapping capacity. A method for removing immunoglobulin dimers by cross-flow filtration using an ultrafiltration membrane (Patent Document 3) has been proposed, but the fractionation performance of immunoglobulin dimers and dimers has been proposed. No data is given. In Patent Document 3, filtration is performed at a very low concentration, and it is estimated that a cake layer capable of separating immunoglobulin monomers and dimers cannot be sufficiently formed.

  In addition, a method is proposed in which biological components having a molecular weight difference of less than 10 times are separated by crossflow filtration while maintaining a level in the range of 5 to 100% of the flux at the transition point (Patent Document 4). However, only the protein separation of 100,000 or less has been reported, and no data on the separation of immunoglobulin monomer and dimer is described.

  In addition, after removing aggregates from an aqueous immunoglobulin solution fractionated from human plasma, it is filtered through a porous polyolefin membrane in the presence of a surfactant having a surfactant activity to reduce anti-complement activity (Patent Document) 5) has been proposed, but there is no specific data on the separation of immunoglobulin monomers and dimers. Furthermore, it is described that it is preferable to use a membrane having a pore size that allows immunoglobulin aggregates (trimers or more) to pass through. It is difficult to separate immunoglobulin monomers and dimers with this membrane. is there.

  As a membrane filtration method for separating immunoglobulin monomer and albumin, there is a method of separating by a method of permeating albumin and blocking immunoglobulin permeation using size separation (Non-patent Document 1). In this method, it is extremely difficult to control in a system using two pumps, and it is difficult to simultaneously remove immunoglobulin dimer and albumin.

Thus, in separation and purification using an ultrafiltration membrane, there is a method of blocking the permeation of both immunoglobulin aggregates and albumins having a size larger than that of immunoglobulin monomers and allowing the immunoglobulin monomers to permeate. There wasn't.
Further, when performing the cross flow filtration method, as in Patent Document 3 and Patent Document 4, it is often performed by constant volume filtration performed by adding the same amount of diluent as the immunoglobulin permeate to the original biological component solution. . However, in this filtration method, since the concentration of the original biological component solution decreases as the filtration proceeds, it may take a long time to increase the recovery rate to 80% or more.
Japanese Patent Publication No.62-3815 JP-A-6-279296 Japanese Patent No. 3746223 Japanese Patent No. 3828143 Japanese Patent Publication No. 7-78025 Bioprocess Technical Sheet No. 1. Nihon Millipore Corporation (1997)

  The present invention uses an ultrafiltration membrane to cross-flow filter an immunoglobulin solution containing at least an immunoglobulin monomer, an aggregate thereof, and a protein having a lower isoelectric point than the immunoglobulin, thereby producing an immunoglobulin monomer. Are provided with high accuracy, an ultrafiltration membrane module thereof, and a crossflow filtration device.

As a result of intensive studies to solve the above-mentioned problems, the present inventors have determined that the molecular weight of the ultrafiltration membrane, the concentration and pH of the immunoglobulin solution, etc. It has been found that the ability to separate immunoglobulin monomers, aggregates and albumin can be expressed without reducing the throughput by optimizing the state.
That is, using an ultrafiltration membrane having a fractional molecular weight of 100,000 or more and less than 500,000, an immunoglobulin solution containing at least an immunoglobulin monomer, an aggregate thereof, and a protein having a lower isoelectric point than immunoglobulin. The immunoglobulin monomer is separated by cross-flow filtration of an immunoglobulin solution characterized in that the immunoglobulin concentration is 1-150 g / L and the pH is equal to or higher than the isoelectric point of the immunoglobulin. And the ultrafiltration membrane module and the crossflow filtration device have been invented.

[化５]

[化６]

[１２]該ポリスルホン系高分子が、ポリビニルピロリドンで親水化されたポリスルホン系高分子であることを特徴とする上記[１０]または[１１]に記載の分離方法。
[１３]該限外濾過膜が、中空糸膜であることを特徴とする上記[１]〜[１２]のいずれかに記載の分離方法。
[１４]下記（イ）〜（ニ）からなる手段の１つ以上の手段を含む装置を用いて行う上記[１]〜[１３]のいずれかに記載の分離方法。
（イ）免疫グロブリン元液の濃度をモニタリングできる手段
（ロ）免疫グロブリン元液の濃度をコントロールできる手段
（ハ）免疫グロブリン元液の線速をコントロールできる手段
（ニ）限外濾過膜の濾過圧力をコントロールできる手段
[１５]上記[１]〜[１４]のいずれかに記載の分離方法に使用するモジュールおよび下記（イ）〜（ニ）からなる手段の１つ以上の手段を含む装置。
（イ）免疫グロブリン元液の濃度をモニタリングできる手段
（ロ）免疫グロブリン元液の濃度をコントロールできる手段
（ハ）免疫グロブリン元液の線速をコントロールできる手段
（ニ）限外濾過膜の濾過圧力をコントロールできる手段
That is, the present invention
[1] An immunoglobulin containing at least a monomer of immunoglobulin, an aggregate thereof, and a protein having a lower isoelectric point than immunoglobulin, using an ultrafiltration membrane having a molecular weight cutoff of 100,000 or more and less than 500,000 Isolating the immunoglobulin monomer by cross-flow filtration of an immunoglobulin solution having an immunoglobulin concentration of 1-150 g / L and a pH equal to or higher than the isoelectric point of the immunoglobulin. Separation method characterized.
[2] The separation method according to [1] above, wherein the aggregate contains at least an immunoglobulin dimer.
[3] The separation method according to [1] or [2], wherein tris (hydroxymethyl) aminomethane is used as a buffer for adjusting the pH of the immunoglobulin solution.
[4] The separation method according to any one of [1] to [3] above, wherein the concentration of the buffer used in the immunoglobulin solution is 0.01 mM to 500 mM.
[5] The separation method according to any one of [1] to [4] above, wherein the pH of the immunoglobulin solution is from the isoelectric point of the immunoglobulin to 11 or less.
[6] The separation method according to any one of [1] to [5] above, wherein the protein having a lower isoelectric point than the immunoglobulin is albumin.
[7] The separation method according to any one of [1] to [6] above, wherein the immunoglobulin concentration is 5 to 100 g / L.
[8] The separation method according to any one of [1] to [7], wherein the cross-flow filtration is performed while maintaining the immunoglobulin concentration substantially constant.
[9] The separation method according to any one of [1] to [8] above, wherein the immunoglobulin is a monoclonal antibody.
[10] The separation method according to any one of [1] to [9], wherein the ultrafiltration membrane is a polysulfone polymer membrane.
[11] The above [10], wherein the polysulfone polymer is at least one or a mixture of two or more polysulfone polymers represented by the following formulas (4) to (6): Separation method.
[Chemical 4]

[Chemical 5]

[Chemical 6]

[12] The separation method according to [10] or [11] above, wherein the polysulfone polymer is a polysulfone polymer hydrophilized with polyvinylpyrrolidone.
[13] The separation method according to any one of [1] to [12], wherein the ultrafiltration membrane is a hollow fiber membrane.
[14] The separation method according to any one of [1] to [13], which is performed using an apparatus including one or more means selected from the following (a) to (d).
(B) Means capable of monitoring the concentration of the original immunoglobulin solution (b) Means capable of controlling the concentration of the immunoglobulin original solution (c) Means capable of controlling the linear velocity of the immunoglobulin original solution (d) Filtration pressure of the ultrafiltration membrane Means to control
[15] An apparatus comprising one or more means selected from the following modules (a) to (d) and a module used in the separation method according to any one of [1] to [14].
(B) Means capable of monitoring the concentration of the original immunoglobulin solution (b) Means capable of controlling the concentration of the immunoglobulin original solution (c) Means capable of controlling the linear velocity of the immunoglobulin original solution (d) Filtration pressure of the ultrafiltration membrane Means to control

By carrying out the separation method of the present invention, high-purity immunity can be obtained by allowing the immunoglobulin monomer to permeate with high permeability and lowering the permeability of aggregates and albumin of immunoglobulin dimer or higher. A globulin monomer can be separated.
The present invention is an extremely simple operation compared to a method such as chromatography, because it is only necessary to adjust to an appropriate solution state and perform cross-flow filtration using an ultrafiltration membrane having a specific fractional molecular weight. Can process large amounts of immunoglobulins. Unlike chemical treatment methods, immunoglobulins are not inactivated or denatured. Further, since clogging does not occur as in known membrane separation, immunoglobulin monomer can be recovered with high permeability, and aggregates and albumin of immunoglobulin dimer or higher can be simultaneously removed.

  Hereinafter, a method for separating an immunoglobulin monomer by an ultrafiltration membrane according to the present invention with high accuracy, an ultrafiltration membrane module thereof, and a crossflow filtration device will be specifically described.

  The immunoglobulin (antibody) according to the present invention is used in the broadest sense, and specifically includes monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, and multispecific antibodies (for example, bispecific antibodies). Etc.

  The immunoglobulin aggregate according to the present invention refers to an immunoglobulin in a state in which the immunoglobulin becomes a dimer or more due to a hydrophobic bond or the like. For example, immunoglobulin dimer, trimer, tetramer, pentamer and the like can be mentioned.

  A monoclonal antibody according to the present invention refers to an antibody obtained from a population of substantially homogeneous antibodies. That is, the individual antibodies that make up the population are the same except for naturally occurring mutations that may be present in small numbers. Monoclonal antibodies are very specific and are directed against a single antigenic site. Furthermore, in contrast to the preparation of conventional polyclonal antibodies, which typically include different antibodies, each monoclonal antibody is directed against a single determinant on the antigen. The modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies. Examples of immunoglobulins (antibodies) according to the present invention include natural human antibodies, humanized antibodies prepared by gene recombination methods, human-type antibodies, fully humanized antibodies, chimeric antibodies, mouse antibodies, and the like.

  Specific examples of antibodies according to the present invention include anti-HER2 receptor antibody, anti-CD20 antibody, anti-IL-8 antibody, anti-VEGF antibody, anti-PSCA antibody, anti-CD11a antibody, anti-IgE antibody, anti-Apo-2 receptor antibody, anti-TNF -Α antibody, anti-tissue factor (TF) antibody, anti-CD3 antibody, anti-CD25 antibody, anti-CD34 antibody, anti-CD40 antibody, anti-tac antibody, anti-CD4 antibody, anti-CD52 antibody, anti-Fc receptor antibody, anti Carcinoembryonic antigen (CEA) antibody, antibody specific for breast epithelial cells, antibody binding to colon cancer cell, anti-CD33 antibody, anti-CD22 antibody, anti-EpCAM antibody, anti-GpIIb / IIIa antibody, anti-RSV antibody, anti-RSV antibody CMV antibody, anti-HIV antibody, anti-hepatitis antibody, anti-αvβ3 antibody, anti-human renal cell carcinoma antibody, anti-human 17-1A antibody, anti-human tube Examples include colorectal tumor antibodies, anti-human melanoma antibodies, anti-human squamous cell carcinoma antibodies, and anti-human leukemia antigen (HLA) antibodies. More specific examples include Muromabab (product name: Orthoclone (OKT3)), Rituximab (product name: Ritaxan), Basiliximab (product name: Simlect), Daclisumab (product name: Zenapax), Palivizumab (product name: Sigma) , Infliximab (product name: Remicade), Gemtuzumab zogamicn (product name: Mylotarg), Alemtuzumab (product name: Mabcampath), Adalimumab (product name: Humira), Omalizumab (product name: Amazumab) Cetuximab (product name: Erbitux) and the like.

  Examples of the molecular target (antigen) of the immunoglobulin (antibody) according to the present invention include, for example, CD protein (eg, CD3, CD4, CD8, CD19, CD20, CD34, and CD40); HER receptor family (eg, EGF receptor, HER2, HER3 or HER4 receptors); cell adhesion molecules (eg LFA-1, Mac1, p150, 95, VLA-4, ICAM-1, VCAM and av / b3 integrins comprising a or b subunits (eg anti-antibodies) -Member of CD11a, anti-CD18 or anti-CD11b antibody); growth factor (eg, VEGF); IgE, blood group antigen; flk2 / flt3 receptor; obesity (OB) receptor; mpl receptor; CTLA-4; Etc. Soluble antigens or fragments bound to other molecules are also molecular targets. For example, in the case of a transmembrane molecule such as a receptor, a fragment of the extracellular region of the receptor is an immune antigen. Alternatively, a cell that expresses a transmembrane molecule may be an immune antigen.

  The protein having a lower isoelectric point than the immunoglobulin according to the present invention refers to a protein having a lower isoelectric point than the immunoglobulin to be separated and purified, and its molecular weight is not particularly related. The isoelectric point of immunoglobulin varies depending on the type, but generally the isoelectric point pI is about 6-9. Therefore, typical examples of proteins having an isoelectric point lower than that of immunoglobulin include albumin (pI4-5), transferrin (pI4-6), and the like.

  In general, cross-flow filtration and dead-end filtration are performed as general-purpose filtration methods as filtration methods. Cross-flow filtration is a process in which a liquid to be treated containing fine particles such as protein is filtered while being supplied to a membrane to separate fine particles having different diameters. It is intended to maintain the fractionation performance by maintaining the state of a stable cake layer over a long period of time while scraping the fine particles (cake layer) deposited on the film surface with the shearing force by the parallel flow of the fine particle solution. . On the other hand, in the dead-end filtration, fine particles are allowed to flow perpendicularly to the membrane surface, so that fine particles accumulate on the membrane surface, the permeation resistance gradually increases with the filtration time, and the permeate fine particle concentration changes. Also called vertical filtration or normal filtration.

  Cross flow filtration according to the present invention (also referred to as cross flow filtration, parallel filtration, or tangential flow filtration) is a method in which an immunoglobulin solution is allowed to flow in parallel to the membrane surface and deposits on the membrane surface by shearing force. This is a filtration system that forms a constant cake layer state in which dynamic equilibrium is established by swirling, and enables continuous operation while maintaining fractionation performance.

  The concentration of the immunoglobulin solution has a great influence on the formation of the cake layer. In order to express the fractionation performance, it is necessary to make the state of an appropriate cake layer. In order to form the cake layer in a short time, it is necessary to consider the lower limit of the concentration. The lower limit concentration of the immunoglobulin solution according to the present invention varies depending on other conditions, but if it is 1 g / L or more, preferably 5 g / L or more, a cake layer that exhibits fractionation performance is formed. On the other hand, the higher the concentration, the thicker the cake layer is formed and the lower the permeability of the immunoglobulin monomer, so it is necessary to consider the upper limit of the concentration at which membrane clogging does not occur. The upper limit concentration of the immunoglobulin solution according to the present invention varies depending on other conditions, but if it is 150 g / L or less, preferably 100 g / L or less, more preferably 50 g / L or less, a sudden membrane occlusion is not caused. It can permeate immunoglobulin monomers. As described above, the lower limit value of the concentration of the immunoglobulin solution is 1 g / L, more preferably 5 g / L, and the upper limit value is 150 g / L, more preferably 100 g / L or less, and even more preferably 50 g / L. It is as follows.

  The concentration change of the immunoglobulin solution during filtration is preferably filtered without changing the conditions in order to maintain the fractionation performance of immunoglobulin monomer and immunoglobulin dimer and the permeation performance of immunoglobulin monomer. . For example, when the concentration of the immunoglobulin solution gradually increases during filtration, filtration clogging may occur and a sufficient amount of permeation may not be obtained. On the other hand, in the constant volume cross flow filtration in which a diluent having the same amount as the permeation amount as in Patent Documents 3 and 4 is added, the concentration of the immunoglobulin solution decreases as the filtration proceeds. In this case, the concentration of the immunoglobulin solution that permeates in the latter half of the filtration decreases, and as a result, a long time is required for recovering in high yield. Therefore, in order to achieve high fractionation performance and permeation performance in a short time, it is preferable to filter the immunoglobulin solution at a constant concentration (constant concentration filtration).

  The change in the concentration of the immunoglobulin solution during filtration is 50 or more, preferably 70 or more, more preferably 80 or more, and 200 or less as the upper limit when the concentration of the immunoglobulin solution before filtration is 100. Preferably it is 150 or less, more preferably 120 or less. In particular, crossflow filtration is most preferred while maintaining the concentration of the immunoglobulin solution substantially constant. Here, “while maintaining the concentration substantially constant” means “while maintaining the concentration with slight fluctuation”. For example, a slight concentration fluctuation that occurs when the concentration is controlled by an operation or an apparatus is included. However, when the remaining amount of the immunoglobulin solution decreases in the latter half of the filtration, and the filtration becomes difficult, in order to recover the immunoglobulin monomer remaining in the membrane or in the apparatus piping, a diluent, buffer solution, water When adding physiological saline and carrying out filtration, it is not necessary to carry out filtration at a constant concentration. In the purification of the immunoglobulin monomer according to the present invention, it is preferable that 50% or more of the immunoglobulin monomer is treated by constant concentration filtration, more preferably 60% or more, and most preferably 70% is constant concentration filtration. It is good to be processed with. For example, after 50% of the immunoglobulin monomer is processed from the start of filtration, it is possible to carry out equal volume filtration until the end, or after performing the equal volume filtration from the start of filtration and processing 20% of the immunoglobulin monomer, After the supply of the diluent is stopped and the solution concentration is returned to the concentration before the start of filtration, constant concentration filtration may be performed. As long as 50% or more of the immunoglobulin monomer is treated by constant concentration filtration by combining constant concentration filtration, equal volume filtration, and dilute-free filtration, the order is not limited. Thereby, an immunoglobulin monomer can be purified in a short time and with a high recovery rate.

The immunoglobulin separation method according to the present invention is a membrane filtration method capable of simultaneously removing immunoglobulin aggregates and proteins having a lower isoelectric point than immunoglobulins. In addition to separating immunoglobulin monomers and their aggregates by pore size in the formed cake layer, proteins with an isoelectric point lower than that of the immunoglobulin are separated from the membrane by electrical repulsion with the formed cake layer. By reducing the permeability, it is believed that as a result, the immunoglobulin monomer of the permeate can be purified with high purity. Therefore, in order to reduce the membrane filterability of proteins having a lower isoelectric point than immunoglobulins, it is necessary to develop an electrical repulsion between the protein having a lower isoelectric point than immunoglobulins and the formed cake layer. The immunoglobulin solution preparation must be strictly performed.
In principle, when purifying an immunoglobulin from an immunoglobulin solution containing a protein having a higher isoelectric point than that of the immunoglobulin, the pH of the immunoglobulin solution can be reduced to a level equal to or lower than that of the immunoglobulin. The mer can be separated.

  The pH of the immunoglobulin solution according to the present invention must be adjusted to be higher than the isoelectric point of the immunoglobulin. By adjusting the pH of the immunoglobulin solution above the isoelectric point of the immunoglobulin, the immunoglobulin is neutrally or weakly negatively charged, and the protein having an isoelectric point lower than that of the immunoglobulin is strongly negatively charged. Therefore, the formed cake layer is also considered to be negatively charged, and a protein having an isoelectric point lower than that of an immunoglobulin is subjected to strong electrical repulsion with the cake layer and hardly permeates through the membrane. The upper limit of the pH of the immunoglobulin solution is not a problem as long as the immunoglobulin does not denature or aggregate, but is generally pH 11 or less, preferably pH 10 or less.

  Buffering agents can be used to adjust the pH of the immunoglobulin solution. The buffering agent according to the present invention is not limited as long as it does not cause denaturation or aggregation of immunoglobulin and can reduce permeation of a protein having a lower isoelectric point than immunoglobulin. For example, tris (hydroxymethyl) aminomethane (Tris), N- [tris (hydroxymethyl) methyl] glycine (Tricine), N, N-bis (2-hydroxyethyl) glycine, N-tris (hydroxymethyl) methyl- 3-aminopropanesulfonic acid (TAPS), 3-[(1,1-dimethyl-2-hydroxyethyl) amino-2-hydroxypropanesulfonic acid] (AMPSO), N-cyclohexyl-2-aminoethanesulfonic acid (CHES) ), N-cyclohexyl-2-hydroxy-3-aminopropanesulfonic acid (CAPSO), 2-amino-2-methyl-1-propanol (AMP), N-cyclohexyl-3-aminopropanesulfonic acid (CAPS), piperazine -1,4-bis (2-ethanesulfonic acid) (P PES) and other good buffers, phosphate buffered saline and other phosphate salts, acetate, glycine, citrate, veronal, borate, succinate, tris (hydroxymethyl) aminomethane, imidazole, etc. Examples include a buffer solution. Among these, a buffering agent using tris (hydroxymethyl) aminomethane (Tris) is particularly preferable.

  The concentration of the buffering agent according to the present invention is not particularly limited as long as it does not cause denaturation or aggregation of immunoglobulin and can reduce the permeation of a protein having a lower isoelectric point than immunoglobulin, but a relatively low concentration is preferable. If the buffer concentration is high, it is presumed that the electrical interaction between the protein and the buffer occurs, the substantial charge on the protein surface is reduced, and the electrical repulsion with the membrane and the cake layer is reduced. Therefore, the lower limit is 0.01 mM or more, preferably 0.1 mM or more, which is a value capable of adjusting the pH of the immunoglobulin solution, and the upper limit is 500 mM or less, preferably 200 mM or less.

  The linear velocity according to the present invention is the velocity of the solution flowing parallel to the film surface. The linear velocity is not particularly limited as long as the cake layer can be expressed without greatly changing the cake layer. For example, the lower limit is 0.1 cm / second or more, preferably 1 cm / second or more. The upper limit is preferably 10 cm / second or more, and the upper limit is 200 cm / second or less, more preferably 100 cm / second or less. If it exceeds 200 cm / second, stress is applied to the immunoglobulin, and aggregation and cloudiness are likely to occur. Conversely, if it is less than 0.1 cm / second, the amount of processing becomes small, resulting in high costs. is there.

  The filtration pressure according to the present invention is not particularly limited as long as the fractionation performance can be expressed. In Patent Document 4, in order to apply a uniform pressure to the entire membrane, a back pressure is applied from the immunoglobulin permeate side, and filtration is performed at a transmembrane pressure (TMP) or less at the transition point to express fractionation performance. I am letting. However, there is a problem that the amount of processing is low due to a device for applying a back pressure and filtration at a low pressure. In the separation method of the present invention, fractionation performance can be maintained even when filtration is performed at a pressure higher than TMP at the transition point, and as a result, the permeation rate of immunoglobulin, that is, the throughput is increased. The filtration pressure according to the present invention has a lower limit of 0.001 MPa or more, preferably 0.005 MPa or more, and an upper limit of 0.30 MPa or less, preferably 0.20 MPa or less. If it is less than 0.001 MPa, the throughput will be low, resulting in poor productivity. On the other hand, if it exceeds 0.30 MPa, a rapid cake layer formation will be caused and there will be a problem that the film will be blocked.

The method for separating an immunoglobulin monomer according to the present invention is a method based on two principles of electrical repulsion by adjusting the size and the solution state. According to the separation based on the size separation, in principle, a biological component having a size of an immunoglobulin dimer or more can also be removed.
The lower limit of the molecular weight of a biological component having a size of an immunoglobulin dimer or more according to the present invention is 300,000 or more, preferably 400,000 or more, more preferably 500,000 or more, and the upper limit is less than 1 million, preferably 900,000. If it is more preferably 800,000 or less, it can be separated from immunoglobulin monomer with high accuracy. If it is less than 300,000, the immunoglobulin monomer cannot be separated with high accuracy. On the other hand, the larger the molecular weight, the higher the accuracy of separation, but if the molecular weight is too high, membrane clogging tends to occur.

Examples of biological components other than immunoglobulin that may be contained in the immunoglobulin solution according to the present invention include proteins, sugar chains, RNA, and DNA. Specific examples include immunoglobulin aggregates, fibrinogen, complex aggregates composed of immunoglobulin and protein A, blood coagulation factors, cell adsorption factors, cell growth factors, enzymes, riboproteins, hormones and aggregates thereof. It is done.
However, it is preferable that it contains a biological component that has an extremely high isoelectric point than immunoglobulins, greatly changes the charge state of the formed cake layer, and increases the permeability of proteins having a lower isoelectric point than immunoglobulins. Absent.

  The above is the case where the immunoglobulin solution according to the present invention contains an immunoglobulin monomer, an aggregate thereof (dimer, trimer,...), And a biological component having a molecular weight of 300,000 to less than 1,000,000. This immunoglobulin solution may be used. In this case, the present invention provides an immunoglobulin 1 amount from an immunoglobulin solution containing an immunoglobulin monomer, an aggregate thereof (dimer, trimer,...) And a biological component having a molecular weight of 300,000 to less than 1,000,000. To separate the body. In addition, as a premise, the present invention naturally means that an immunoglobulin monomer is separated from an immunoglobulin solution which does not contain or hardly contains the above-mentioned biological component.

The immunoglobulin solution according to the present invention may contain a virus.
Examples of the virus according to the present invention include parvovirus and poliovirus having a diameter of about 18 to 24 nm, Japanese encephalitis virus and hepatitis virus having a diameter of about 40 to 45 nm, and HIV having a diameter of about 80 to 100 nm. The one that most needs to be removed is the parvovirus. Since parvovirus is the smallest virus that has been confirmed at present, in the case of size fractionation by membrane, if parvovirus can be removed, other large viruses can be removed.

Since viruses cause serious infections such as hepatitis, acquired immune deficiency, Japanese encephalitis, and childhood paralysis, the virus log removal rate (LRV) is preferably 4 or more. LRV is calculated by the following formula (7).
LRV = log [(No × Vo) / (Nf × Vf)] (7)
LRV: Virus logarithm removal rate No: Virus infectivity titer in the original immunoglobulin solution before filtration (TICD ₅₀ / mL)
Nf: virus infectivity in the immunoglobulin permeate (TICD ₅₀ / mL)
Vo: Volume of the original immunoglobulin solution before filtration (mL)
Vf: Volume of immunoglobulin permeate (mL)
Examples of the parvovirus used for the evaluation of virus removability include human parvovirus (B19), porcine parvovirus, canine parvovirus, feline parvovirus, Minute of rice, and the like. These viruses have a diameter of about 18 to 24 nm and can be evaluated using any of them. However, it is preferable to use porcine parvovirus or canine parvovirus for which a simple infectivity evaluation method has been established. .

The “ultrafiltration membrane” according to the present invention is a membrane used for separating immunoglobulin monomers.
As an ultrafiltration membrane, a membrane having a desired fractional molecular weight can be produced and is not limited at all unless immunoglobulin is adsorbed. For example, a polysulfone-based polymer membrane, an aromatic ether-based polymer membrane, a fluorine-based high-molecular membrane is used. Examples thereof include a molecular film, an olefin polymer film, a cellulose film, a (meth) acrylic polymer film, a (meth) acrylonitrile polymer film, and a vinyl alcohol polymer film. A polysulfone-based polymer membrane is preferable.

[化９]

[化１０]

これらの高分子を二種以上、組み合わせて実施することも可能である。ポリスルホン系高分子は、必要に応じて高分子末端および／または主鎖中にエステル化、エーテル化、エポキシ化など各種変性を実施することができる。また、免疫グロブリンの静電気的な特性との相性から、アミノ基、モノアルキルアミノ基、ジアルキルアミノ基、カルボキシル基、スルフォニル基、スルホン酸基などの化学構造を必要に応じて導入しても良い。 The polysulfone polymer membrane according to the present invention is not particularly limited, and any polymer having a sulfone group in the molecule can be used. Examples of the polysulfone polymer include polysulfone represented by the following formula (8), polyethersulfone represented by the following formula (9), polyarylsulfone represented by the following formula (10), and the like. . In the formula, l, m, and n represent repeating units.

[Chemical 8]

[Chemical 9]

[Chemical Formula 10]

It is also possible to carry out by combining two or more of these polymers. The polysulfone polymer can be subjected to various modifications such as esterification, etherification, and epoxidation in the polymer terminal and / or main chain as necessary. Moreover, chemical structures such as an amino group, a monoalkylamino group, a dialkylamino group, a carboxyl group, a sulfonyl group, and a sulfonic acid group may be introduced as necessary in view of compatibility with the electrostatic properties of immunoglobulins.

  The weight average molecular weight of the polysulfone polymer according to the present invention is preferably 5,000 or more, preferably 10,000 or more, more preferably 20,000 or more as a lower limit, and 1 million or less, preferably as an upper limit. It is good to use 500,000 or less, more preferably 300,000 or less. Within this range, sufficient strength and film formability can be obtained.

  In the present invention, it is preferable to use a hydrophilic polymer together with the polysulfone-based polymer in order to control the pore size of the membrane and to impart hydrophilicity. It improves the contact between the immunoglobulin solution subjected to separation treatment by hydrophilization and the ultrafiltration membrane comprising the polysulfone polymer of the present invention.

  The kind of the hydrophilic polymer that imparts hydrophilicity to the polysulfone polymer according to the present invention is not particularly limited. For example, polyvinylpyrrolidone, polyethylene glycol, polypropylene glycol, polyethylene oxide, polyethylene glycol-polypropylene glycol block copolymer, polyvinyl alcohol, polyacrylamide, poly-N, N-dimethylacrylamide, poly-N-isopropylacrylamide, polyhydroxyacrylate, poly Examples include hydroxy methacrylate, carboxymethyl cellulose, starch, corn starch, polychitosan, and polychitin. Among these, polyvinyl pyrrolidone has good compatibility with the polysulfone polymer, and is particularly preferable for enhancing the hydrophilicity of the entire membrane.

  The weight average molecular weight of the hydrophilic polymer imparting hydrophilicity to the polysulfone polymer membrane according to the present invention is 1,000 or more, preferably 5,000 or more as a lower limit, and 2 million or less, preferably as an upper limit. Is preferably 1.2 million or less. For example, various grades of polyvinyl pyrrolidone are commercially available from BASF and have a weight average molecular weight of 9,000 (K17), and similarly 45,000 (K30), 450,000 (K60), 900,000. (K80) and 1,200,000 (K90) are preferably used, and in order to obtain the intended use, characteristics, and structure, each may be used alone or in combination of two or more. . In the present invention, it is most preferable to use K90 alone.

  The content of the hydrophilic polymer in the polysulfone polymer membrane according to the present invention is not particularly limited as long as the immunoglobulin is not adsorbed on the membrane. For example, the lower limit is 0.1% by weight or more, preferably 0.3% by weight or more, more preferably 0.5% by weight or more. The upper limit is 10% by weight or less, preferably 8% by weight or less, and more preferably. Is preferably 5% by weight or less.

The molecular weight cut off of the polysulfone-based polymer membrane according to the present invention is only required to sufficiently separate the immunoglobulin monomer and the dimer, and the lower limit is 100,000 or more, preferably 150,000 or more, more preferably 250,000 or more. The upper limit should be set to less than 500,000, preferably 450,000 or less, and more preferably 400,000 or less. If the molecular weight cut-off is less than 100,000, there is a problem that the amount of immunoglobulin permeated decreases, and if it is more than 500,000, both the immunoglobulin monomer and dimer permeate through the membrane. Decreases.
The molecular weight cut-off according to the present invention includes proteins such as albumin (66,000), γ-globulin (160,000), catalase (232,000), ferritin (440,000), thyroglobulin (669,000), and polyethylene. Dead end filtration is performed using glycol (PEG), dextran, or the like, and the molecular weight is calculated as the molecular weight at which the rejection is 90% from the relationship between the molecular weight and the rejection.

  The method for producing the polysulfone-based polymer membrane according to the present invention is not limited in any way, and examples thereof include a wet film-forming method. The wet film formation method is a method in which a membrane stock solution obtained by dissolving a membrane material in a good solvent is brought into contact with a coagulation liquid composed of another solvent that is miscible with the good solvent in the membrane stock solution but is not compatible with the membrane material. In this method, concentration-induced phase separation is generated from the contact surface to obtain a membrane.

  The membrane stock solution used in the wet film forming method for obtaining the polymer membrane according to the present invention is not limited as long as a polysulfone polymer having the desired structure and performance can be produced. For example, the entire membrane stock solution is 100% by weight. In this case, the lower limit of the concentration range of the polysulfone polymer is 1% by weight or more, preferably 2% by weight or more, and particularly preferably 3% by weight or more. Moreover, as an upper limit, the solution which melt | dissolved uniformly in 45 weight% or less, Preferably it is 35 weight% or less, Most preferably, it is 25 weight% or less is used suitably. The hydrophilic polymer has a lower limit of 0.1% by weight or more, preferably 0.5% by weight or more, and an upper limit of 20% by weight or less, preferably 10% by weight or less, and a uniformly dissolved solution is preferably used. . In addition, the lower limit of the temperature of the membrane stock solution is preferably 0 ° C. or higher, preferably 10 ° C. or higher, particularly preferably 25 ° C. or higher, and the upper limit is a good solvent boiling point or lower in the membrane stock solution. If it is this temperature condition, the viscosity suitable for processing into a film | membrane preferable as a film | membrane stock solution can be obtained.

  The good solvent used in the wet film-forming method for obtaining the polysulfone polymer film according to the present invention can dissolve 10 g or more in 100 g pure water at 20 ° C., and dissolves 5% by weight or more of the polysulfone polymer of the membrane material. However, there is no limitation as long as it is miscible with water. Specifically, N-methyl-2-pyrrolidone, N, N-dimethylacetamide, dimethyl sulfoxide, N, N-dimethylformamide And γ-butyrolactone. These can be used in combination of two or more.

  The coagulating liquid used in the wet film forming method for obtaining the polysulfone-based polymer film according to the present invention is a substance that can cause concentration-induced phase separation when in contact with the membrane stock solution and can form a film from the contact surface. There is no limitation. For example, pure water, a monoalcohol solvent, a polyol solvent, or a mixture of two or more of these is preferably used. Examples of the monoalcohol solvent include methanol, ethanol, propanol and the like. Examples of the polyol solvent include ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, glycerin, propylene glycol, 1,2-butanediol, 1,4-butanediol, and the like. Polyvinyl alcohol, polyethylene glycol, polypropylene glycol, polyethylene oxide, polyethylene glycol-polypropylene glycol block copolymer, polyacrylamide, polyvinyl pyrrolidone, polyhydroxy acrylate, polyhydroxy methacrylate, polyacrylic acid, polymethacrylic acid, polyitaconic acid in the coagulation liquid Water-soluble polymers such as polyfumaric acid, polycitraconic acid, poly-p-styrenesulfonic acid, sodium poly-p-styrenesulfonate, N, N-dimethylacrylamide, carboxymethylcellulose, starch, corn starch, polychitosan, and polychitin It is also possible to add. Although it depends on the molecular weight and the amount of the water-soluble polymer to be added, the filtration performance can be improved by adding these.

  The coagulation liquid may contain a good solvent such as N-methyl-2-pyrrolidone, N, N-dimethylacetamide, dimethyl sulfoxide, N, N-dimethylformamide, and γ-butyrolactone. In particular, when using a coagulation liquid containing a good solvent in a non-solvent, the composition differs depending on the composition of the membrane stock solution, the contact temperature between the membrane stock solution and the coagulation liquid, etc. In this case, 90% by weight or less is preferable as the weight% of the good solvent. Within this range, concentration-induced phase separation necessary and sufficient for forming a film can be sufficiently achieved.

  The film forming temperature in the wet film forming method according to the present invention is a temperature at which the membrane stock solution and the coagulating liquid are brought into contact with each other to cause concentration-induced phase separation. It depends on the temperature. The lower limit of the film formation temperature is 0 ° C. or higher, preferably 10 ° C. or higher, particularly preferably 25 ° C. or higher. The upper limit is a temperature below each boiling point of the membrane stock solution or coagulation liquid, preferably a temperature lower by 5 ° C. or more from each boiling point, particularly preferably a temperature lower by 10 ° C. or more from the boiling point. For hollow fiber membranes, it depends on the temperature of the double nozzle. In a flat membrane, it is determined by the coagulation liquid temperature.

  The membrane stock solution used in the wet film formation method for obtaining the polysulfone-based polymer membrane according to the present invention, the coagulation solution, especially the coagulation solution that passes through the inside of the yarn when producing the hollow fiber membrane (hereinafter referred to as the internal coagulation solution) is dissolved after uniform dissolution. It is desirable to remove the gas. By removing the dissolved gas, film defects due to foaming of the dissolved gas can be remarkably improved. Further, by removing oxygen from the dissolved gas, the oxidation reaction to the material due to film processing at a high temperature is reduced. This step may be omitted when no gas is dissolved in the membrane stock solution, the coagulation solution, and the internal coagulation solution.

When producing a hollow fiber membrane by the wet film-forming method according to the present invention, in order to further promote the coagulation by the membrane stock solution and the internal coagulation solution discharged from the double spinning nozzle, It can be provided and brought into contact with a coagulation liquid (hereinafter referred to as an external coagulation liquid) filled in the coagulation tank.
When producing a hollow fiber membrane by the wet film-forming method of the present invention, the cross-sectional structure of the hollow fiber membrane is not only a uniform structure but also various non-uniform structures. And the temperature and humidity of the space from the spinneret to the external coagulation liquid surface can be adjusted. Although there is no limitation as long as the temperature and humidity of the space can be adjusted, for example, the lower limit of the free running distance is 0.001 m or more, preferably 0.005 m or more, particularly preferably 0.01 m or more, and the upper limit is 2.0 m or less, preferably Is 1.5 m or less, particularly preferably 1.2 m or less. The temperature in the space from the spinneret to the external solidification surface is 10 ° C. or higher, preferably 20 ° C. or higher, particularly preferably 25 ° C. or higher as the lower limit. The humidity varies depending on the temperature, but the lower limit is 0% or more, preferably 10% or more, particularly preferably 30% or more, and the upper limit is 100% or less.

  The winding speed in the case of producing a hollow fiber membrane by the wet film-forming method according to the present invention is as follows: various factors as production conditions, the shape of the nozzle, the composition of the spinning stock solution, the composition of the internal coagulating liquid and the external coagulating liquid, and the stock solution Although the speed may vary depending on the temperature of each coagulating liquid, a speed of approximately 300 m / hour to 9,000 m / hour is selected.

In the wet film-forming method according to the present invention, after the solidification by the coagulating liquid, the solvent removal can be promoted by immersing in a solvent removal tank in order to increase the strength of the film. The solvent removal liquid is a solvent that can remove the remaining solvent after concentration-induced phase separation by the coagulation liquid, and any solvent that does not dissolve the membrane can be used. In general, water, ethanol or the like is often used.
An undried polysulfone-based polymer membrane obtained by a wet film-forming method is dried at a temperature at which no membrane breakage occurs during drying. For example, the lower limit of the drying temperature is 20 ° C. or higher, preferably 30 ° C. or higher, more preferably 50 ° C. or higher, and the upper limit is the melting temperature or lower, preferably 150 ° C. or lower, more preferably 140 ° C. or lower. The time required for drying is determined by the relationship with the drying temperature, but is generally selected from 0.01 hours to 48 hours.

（Ｒ^１、Ｒ^２、Ｒ^３、Ｒ^４、Ｒ^５、Ｒ^６は水素、炭素数１以上６以下を含む有機官能基、または、酸素、窒素または珪素を含有する炭素数６以下の非プロトン性有機官能基であり、それぞれ同一であっても、異なっても構わない。構造式中のｑは繰り返し単位数である。異なる繰り返し単位を２成分以上含む共重合体でも構わない。） The aromatic ether polymer film according to the present invention is not particularly limited, and examples of the aromatic ether polymer film include those represented by the following formula (11).

[Chemical 11]

(R ¹ , R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are hydrogen, an organic functional group containing 1 to 6 carbon atoms, or an aprotic having 6 or less carbon atoms containing oxygen, nitrogen or silicon. The organic functional groups may be the same or different, and q in the structural formula represents the number of repeating units, and may be a copolymer containing two or more different repeating units.)

The terminal phenolic hydroxyl group of the aromatic ether polymer according to the present invention is esterified, etherified, epoxidized, etc. as necessary in order to maintain a pH that can stably exist in an immunoglobulin solution. Various modifications can be performed. In addition, due to compatibility with the electrostatic properties of immunoglobulins, chemical structures such as amino groups, monoalkylamino groups, dialkylamino groups, carboxyl groups, sulfonyl groups, and sulfonic acid groups can be introduced into the polymer terminals as needed. .
The aromatic ether polymer film according to the present invention is mainly composed of an aromatic ether polymer, but contains other high molecular weight substances and additives as long as the characteristics of the aromatic ether polymer are not impaired. You may do it. It is also possible to carry out by combining two or more of these polymers. For example, polystyrene or a derivative thereof may be contained.

  The weight average molecular weight of the aromatic ether polymer according to the present invention is 5,000 or more, preferably 10,000 or more, particularly preferably 20,000 or more as the lower limit, and 1,000,000 or less, preferably 500,000 or less as the upper limit. Particularly preferred is 300,000 or less. Within this range, sufficient strength and film formability can be obtained.

In the present invention, it is preferable to use a hydrophilic polymer together with the aromatic ether polymer in order to control the pore size of the membrane and to impart hydrophilicity. The contact between the immunoglobulin solution subjected to the separation treatment by hydrophilization and the ultrafiltration membrane made of the aromatic ether polymer of the present invention is improved.
The hydrophilic polymer according to the present invention is not limited as long as it can impart hydrophilicity. In order to reduce the electrical interaction with the immunoglobulin, it is desirable to be nonionic without a charged structure.

The hydrophilic polymer according to the present invention may be any polymer compound as long as it can impart hydrophilicity.
Examples of the hydrophilic polymer include polyethylene glycol, polypropylene glycol, polyethylene oxide, polyethylene glycol-polypropylene glycol block copolymer, polyvinyl alcohol, polyacrylamide, poly-N, N-dimethylacrylamide, poly-N-isopropylacrylamide, Examples thereof include hydrophilic polymer compounds such as polyvinylpyrrolidone, polyhydroxyacrylate, polyhydroxymethacrylate, carboxymethylcellulose, starch, corn starch, polychitosan, and polychitin. In addition, surfactants, block copolymers, and graft copolymers containing these substances as hydrophilic segments and hydrophobic segments can be sufficiently utilized as hydrophilic polymers. For example, a polystyrene-polyethylene glycol block copolymer is preferable.

  Since the polystyrene-polyethylene glycol block copolymer according to the present invention has polyethylene glycol having high hydrophilicity in the hydrophilic segment, it can be effectively used as a hydrophilic polymer. Moreover, these can also be used in combination of 2 or more types. Among these, polyethylene glycol and block copolymers and graft copolymers containing polyethylene glycol as hydrophilic segments can be preferably used. Among them, polystyrene-polyethylene glycol block copolymers are particularly aromatic ethers. It can be suitably used as a hydrophilic polymer that improves the hydrophilicity of the polymer film.

  The molecular weight of the hydrophilic polymer according to the present invention is appropriately selected depending on the production method and its conditions. For example, when the film formation method is a wet film formation method and a non-halogen water-soluble organic solvent is used as the solvent, the solubility of the aromatic ether polymer having high solvent resistance is extremely low. Therefore, when the hydrophilic polymer is blended with the membrane stock solution, it is necessary to appropriately select the molecular weight and the addition amount of the hydrophilic polymer in order to obtain a uniformly dissolved membrane stock solution. In order to use a sufficient amount of hydrophilic polymer, the number average molecular weight of the hydrophilic polymer is preferably 300 or more and 100,000 or less, for example. If it is this area | region, it can melt | dissolve enough in the good solvent used for film-forming. A more preferable lower limit is 400 or more, and a particularly preferable lower limit is 500 or more. More preferably, the upper limit is 70,000 or less, and particularly preferably 50,000 or less.

  When the hydrophilic polymer according to the present invention is a compound comprising a hydrophobic segment and a hydrophilic segment, the number average molecular weight of the hydrophilic segment of the hydrophilic polymer is preferably 300 or more and 100,000 or less. If it is this area | region, it can melt | dissolve enough in the good solvent used for film-forming. A more preferable lower limit is 400 or more, and a particularly preferable lower limit is 500 or more. More preferably, the upper limit is 70,000 or less, and particularly preferably 50,000 or less.

（Ｒ^７、Ｒ^８、Ｒ^９、Ｒ^１０、Ｒ^１１、Ｒ^１２、Ｒ^１３、Ｒ^１４は水素、フッ素を除くハロゲン原子、炭素数１以上６以下を含む有機官能基、または、酸素、窒素または珪素を含有する炭素数６以下の官能基であり、それぞれ同一であっても、異なっても構わない。構造式中のｓは繰り返し単位数である。構造範囲内で異なる繰り返し単位を２成分以上含む共重合体でも構わない。） The polystyrene-polyethylene glycol block copolymer according to the present invention is a block copolymer comprising a segment derived from a polystyrene polymer and a segment derived from a polyethylene glycol polymer.
As the polystyrene polymer forming the segment derived from the polystyrene polymer of the polystyrene-polyethylene glycol block copolymer used in the present invention, a polystyrene polymer comprising a repeating unit represented by the following formula (12) is preferable.

[Chemical 12]

(R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ , R ¹⁴ are hydrogen, a halogen atom excluding fluorine, an organic functional group containing 1 to 6 carbon atoms, or oxygen, nitrogen Or a silicon-containing functional group having 6 or less carbon atoms, which may be the same or different, and s in the structural formula is the number of repeating units. It may be a copolymer including the above.)

  The number average molecular weight of the segment derived from the polystyrene-based polymer of the polystyrene-polyethylene glycol block copolymer according to the present invention needs to be 300 or more and 1,000,000 or less. If it is this area | region, it can fully melt | dissolve in the good solvent used for film-forming, and it can reduce the elution property with respect to aqueous solution. The more preferable lower limit is 500 or more, and the particularly preferable lower limit is 700 or more. The upper limit is more preferably 500,000 or less, and the particularly preferable upper limit is 300,000 or less.

[化１４]

（Ｒ^１５は、炭素数３以上、３０未満の有機官能基である。特に親水性が大きく低下させることがなければ、Ｒ^１５にエーテル基、エステル基、水酸基、ケトン基、カルボン酸基を含有しても構わない。ｔおよびｕは繰り返し単位数である。） The polyethylene glycol polymer forming a segment derived from the polyethylene glycol polymer of the polystyrene-polyethylene glycol block copolymer used in the present invention is a repeating unit represented by the following formula (13) and / or (14). A polyethylene glycol polymer is preferable.

[Chemical 13]

[Chemical 14]

(R ¹⁵ is an organic functional group having 3 or more carbon atoms and less than 30. Unless the hydrophilicity is greatly lowered, R ¹⁵ contains an ether group, an ester group, a hydroxyl group, a ketone group, or a carboxylic acid group. (T and u are the number of repeating units.)

  The number average molecular weight of the segment derived from the polyethylene glycol polymer of the polystyrene-polyethylene glycol block copolymer used in the present invention needs to be, for example, 300 or more and 100,000 or less. If it is this area | region, sufficient hydrophilic property will be obtained at that time if it can melt | dissolve sufficiently in the good solvent used for film-forming. A more preferable lower limit is 400 or more, and a particularly preferable lower limit is 500 or more. More preferably, the upper limit is 70,000 or less, and particularly preferably 50,000 or less.

  As the composition ratio of the segment derived from the polystyrene polymer of the polystyrene-polyethylene glycol block copolymer according to the present invention and the segment derived from the polyethylene glycol polymer, the segment derived from the polystyrene polymer is all polystyrene-polyethylene. It is necessary that the content is 10% by weight or more and 99% by weight or less of the glycol block copolymer. In this composition ratio, sufficient hydrophilicity can be expressed and elution is suppressed. A more preferred lower limit is 20% by weight or more, a particularly preferred lower limit is 30% by weight or more, a more preferred upper limit is 98% by weight or less, and a particularly preferred upper limit is 97% by weight or less.

The block structure of the polystyrene-polyethylene glycol block copolymer according to the present invention is a diblock copolymer composed of two segments, a triblock copolymer composed of three segments, and four or more It may be a multi-block copolymer composed of segments. Moreover, you may be a mixture of these 2 or more types of block copolymers. The number average molecular weights of the respective segments to be constructed may be the same or different.
In the polystyrene-polyethylene glycol block copolymer according to the present invention, the segment derived from the polystyrene polymer and the segment derived from the polyethylene glycol polymer need to be directly chemically bonded at the polymer terminal portion. For production, if necessary, a low molecular compound and / or an organic functional group may be used as a spacer for connecting the segment derived from the polystyrene polymer and the segment derived from the polyethylene glycol polymer. . When the number average molecular weight of the low molecular weight compound and / or the organic functional group is 500 or less, the low molecular weight compound and / or the organic functional group can be expressed without reducing the effect of the segment derived from the polystyrene polymer and the segment derived from the polyethylene glycol polymer. Specific examples include low molecular weight compounds between polystyrene and polyethylene glycol formed when polyethylene glycol is condensed after polymerizing styrene using a radical polymerization initiator having a reactive functional group.

  An example of a method for producing the polystyrene-polyethylene glycol block copolymer according to the present invention is a method using a radical polymerization initiator having a reactive functional group. Specifically, an azo radical polymerization initiator having a carboxylic acid group is used, and after chemically converting the carboxylic acid group to an acid chloride group, the terminal has an acid chloride group by radical polymerization of styrene. Polystyrene is obtained. Subsequently, a polystyrene-polyethylene glycol block copolymer can be obtained by condensation with polyethylene glycol (Polymer Papers, 1976, Vol. 33, P131). A polystyrene-polyethylene glycol block copolymer can also be obtained by radical polymerization of styrene using a polyethylene glycol unit-containing polymer azo polymerization initiator. Another example of the synthesis method is a method using living polymerization. Specifically, styrene is polymerized using living radical polymerization with a nitroxide compound, and a polymer in which the nitroxide compound is bonded to the polymer terminal can be obtained. A polymer end can be converted into a hydroxyl group by hydrolysis, and a polystyrene-polyethylene glycol block copolymer can be obtained by a coupling reaction with polyethylene glycol (Polymer, 1998, Vol. 39, No. 4, P911). .

  The method of hydrophilizing an ultrafiltration membrane composed of an aromatic ether polymer using a hydrophilic polymer in the present invention is, for example, a blend method in which a hydrophilic polymer is mixed in advance at the time of film formation, Examples of the method include a coating method in which a film is immersed in a solution, and then dried to leave a hydrophilic polymer, and a method in which a hydrophilic acrylic monomer, a methacrylic monomer, an acrylamide monomer, or the like is graft-polymerized on the film surface. It is also possible to perform a combination of two or more of these methods. A blending method or a coating method in which the aromatic ether polymer is not chemically modified is preferable, and a blending method capable of performing a hydrophilization treatment in a single step is particularly preferable in terms of production.

The molecular weight cut off of the aromatic ether polymer membrane according to the present invention is only required to sufficiently separate immunoglobulin monomers and dimers, and the lower limit is 100,000 or more, preferably 150,000 or more, more preferably 250,000. The upper limit is good, and the upper limit should be set to less than 500,000, preferably 450,000 or less, more preferably 400,000 or less. If the molecular weight cut-off is less than 100,000, there is a problem that the amount of immunoglobulin permeated decreases, and if it is more than 500,000, both the immunoglobulin monomer and dimer permeate through the membrane. Decreases.
The molecular weight cut-off according to the present invention includes proteins such as albumin (66,000), γ-globulin (160,000), catalase (232,000), ferritin (440,000), thyroglobulin (669,000), and PEG. Then, dead end filtration is performed using dextran or the like, and the molecular weight is calculated as the molecular weight at which the rejection is 90% from the relationship between the molecular weight and the rejection.

The method for producing the aromatic ether polymer film according to the present invention is not limited in any way, and examples thereof include a wet film forming method. The wet film formation method is to bring a membrane stock solution in which a membrane material is dissolved in a good solvent into contact with a coagulation liquid composed of another solvent that is miscible with the good solvent in the membrane stock solution but is incompatible with the membrane material. In this method, concentration-induced phase separation is generated from the contact surface to obtain a membrane.
The good solvent used in the wet film formation method for obtaining the aromatic ether polymer film according to the present invention is one that can stably dissolve 5% by weight or more of the aromatic ether polymer as the film material under the film formation conditions. Any solvent can be used. However, it is preferable to use a non-halogen water-soluble organic solvent from the viewpoint of environment and cost. The water-soluble organic solvent in the present invention refers to a solvent that can be dissolved in 10 g or more in 100 g pure water at 20 ° C., and more preferably is miscible with water. Specific examples include N-methyl-2-pyrrolidone, N, N-dimethylacetamide, dimethyl sulfoxide, N, N-dimethylformamide, and γ-butyrolactone. These can be used in combination of two or more.

As an example of the membrane stock solution used in the wet film-forming method for obtaining the aromatic ether polymer membrane according to the present invention, 0.1 wt% or more, preferably 0.5 wt. %, Particularly preferably 1% by weight or more, and an upper limit of 45% by weight or less, preferably 35% by weight or less, particularly preferably 25% by weight or less, and a uniformly dissolved solution is preferably used.
Further, when the total membrane stock solution is 100% by weight, the lower limit of the use range of the aromatic ether polymer is 1% by weight or more, preferably 2% by weight or more, and particularly preferably 3% by weight or more. Moreover, as an upper limit, the solution which melt | dissolved uniformly in 45 weight% or less, Preferably it is 35 weight% or less, Most preferably, it is 25 weight% or less is used suitably.
In addition, the lower limit of the temperature of the membrane stock solution is preferably 25 ° C. or higher, preferably 65 ° C. or higher, particularly preferably 80 ° C. or higher, and the upper limit is a good solvent boiling point or lower in the membrane stock solution. By using this temperature condition, the solubility of the aromatic ether polymer can be increased, and a viscosity suitable for processing into a film preferable as a film stock solution can be obtained.

（Ｒ^１６は炭素数１以上、２０以下を含む有機官能基、または、酸素原子を１つ以上と炭素数１以上、２０以下とを含む構造であり、Ｒ^１６に水酸基、エーテル結合、エステル基、ケトン基、カルボン酸基などを１つ以上含んでいてもよい。）
式（１５）に表されるポリオール系溶媒の一例として、エチレングリコール、ジエチレングリコール、トリエチレングリコール、テトラエチレングリコール、グリセリン、プロピレングリコール、１，２−ブタンジオール、１，４−ブタンジオールなどが挙げられる。 The coagulating liquid used in the wet film forming method for obtaining the hydrophilic aromatic ether polymer film according to the present invention can cause concentration-induced phase separation when in contact with the film stock solution, and can form a film from the contact surface. A substance. Specifically, pure water, a monoalcohol solvent, a polyol solvent represented by the following formula (15), or a mixture of two or more of these are preferably used.

[Chemical 15]

(R ¹⁶ is the number 1 or more carbon atoms, 20 an organic functional group include the following, or an oxygen atom and 1 or more and having 1 or more carbon atoms, a structure including a 20 or less, hydroxyl groups in R ^16, an ether bond, an ester group , One or more ketone groups, carboxylic acid groups, and the like.
Examples of the polyol solvent represented by the formula (15) include ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, glycerin, propylene glycol, 1,2-butanediol, 1,4-butanediol, and the like. .

  The water permeation performance can be controlled by the viscosity of the coagulation liquid used in the wet film forming method for obtaining the aromatic ether polymer film. It has been found that by increasing the viscosity of the coagulation liquid, the permeation of the coagulation liquid into the stock solution becomes gradual, and as a result, the water permeability of the produced membrane is improved. In order to obtain high water permeability, the viscosity of the coagulation liquid is preferably 3 cp or more at 20 ° C. More preferably, it is 5 cp or more. Polyvinyl alcohol, polyethylene glycol, polypropylene glycol, polyethylene oxide, polyethylene glycol-polypropylene glycol block copolymer, polyacrylamide, polyvinyl pyrrolidone, polyhydroxy acrylate, polyhydroxy methacrylate, polyacrylic acid, polymethacrylic acid to increase the viscosity of the coagulation liquid Water such as acid, polyitaconic acid, polyfumaric acid, polycitraconic acid, poly-p-styrene sulfonic acid, sodium poly-p-styrene sulfonate, N, N-dimethylacrylamide, carboxymethyl cellulose, starch, corn starch, polychitosan, polychitin It is possible to add a functional polymer. It is also preferable to contain a high viscosity solvent of 5 cp or more at 20 ° C. such as the above-mentioned polyol solvent.

The viscosity of the coagulating liquid according to the present invention is a value measured using a glass capillary viscometer. As a measuring method, the coagulating liquid is put into a glass capillary viscometer made constant in a 20 ° C. constant temperature water bath, and 30 The kinematic viscosity can be obtained by allowing the sample to stand for more than a minute and then measuring that it has reached a constant temperature. From the obtained kinematic viscosity value, the viscosity of the coagulation liquid can be obtained by the following formula (16). In addition, as a glass capillary viscometer, an Uberote viscometer manufactured by Shibata Kagaku Co., Ltd. can be used.
ν = η / ρ (16)
([Nu: kinematic viscosity ^{(mm 2 / s), η} : viscosity (cp), ρ: Density (g / ^cm 3))

  In a coagulation liquid used in a wet film forming method for obtaining an aromatic ether polymer film according to the present invention, N-methyl-2-pyrrolidone, N, N-dimethylacetamide, dimethyl sulfoxide, N, N-dimethylformamide, It is also possible to include a good solvent such as γ-butyrolactone. In particular, when using a coagulation liquid containing a good solvent in a non-solvent, the composition differs depending on the composition of the membrane stock solution, the contact temperature between the membrane stock solution and the coagulation liquid, etc. In this case, the lower limit of 0% by weight and the upper limit of 90% by weight are preferable as the weight% of the good solvent. Within this range, concentration-induced phase separation necessary and sufficient for forming a film can be sufficiently achieved.

  The film forming temperature in the wet film forming method according to the present invention is a temperature at which the membrane stock solution and the coagulating liquid are brought into contact with each other to cause concentration-induced phase separation. It depends on the temperature. In the case of a flat membrane, it is determined by the coagulation liquid temperature. The lower limit of the film formation temperature is 25 ° C. or higher, preferably 80 ° C. or higher, particularly preferably 90 ° C. or higher. The upper limit is a temperature below each boiling point of the membrane stock solution or coagulation liquid, preferably a temperature lower by 5 ° C. or more from each boiling point, particularly preferably a temperature lower by 10 ° C. or more from the boiling point. In particular, when the film stock solution and the coagulating liquid are in contact with each other within a film forming temperature of 80 ° C. or higher and a temperature range of 10 ° C. or lower from each boiling point, a particularly high strength film can be obtained.

  The membrane stock solution used in the wet film forming method for obtaining the aromatic ether polymer membrane according to the present invention, the coagulating solution, especially the coagulating solution passing through the inside of the yarn during the production of the hollow fiber membrane (hereinafter referred to as the internal coagulating solution) is uniformly dissolved. It is desirable to remove dissolved gas. By removing the dissolved gas, film defects due to foaming of the dissolved gas can be remarkably improved. Further, by removing oxygen from the dissolved gas, the oxidation reaction to the material due to film processing at a high temperature is reduced.

When producing a hollow fiber membrane by the wet film-forming method according to the present invention, in order to further accelerate the coagulation by the membrane stock solution and the internal coagulation liquid that has come out of the double spinning nozzle, a tank (hereinafter referred to as a coagulation tank) immediately below the spinning nozzle. And can be brought into contact with a coagulating liquid (hereinafter referred to as an external coagulating liquid) filled in the coagulating tank.
When producing a hollow fiber membrane by the wet film-forming method according to the present invention, the cross-sectional structure of the hollow fiber membrane is not only a uniform structure but also various non-uniform structures. The distance to the surface (hereinafter referred to as idle running distance) and the temperature and humidity of the space from the spinneret to the external coagulation liquid surface can be adjusted.
When producing a hollow fiber membrane by the wet film-forming method according to the present invention, the lower limit of the free running distance is 0.01 m or more, preferably 0.05 m or more, particularly preferably 0.1 m or more, and the upper limit is 2.0 m or less. , Preferably 1.5 m or less, particularly preferably 1.2 m or less. The temperature in the space from the spinneret to the external solidification surface is 20 ° C. or higher, preferably 50 ° C. or higher, particularly preferably 80 ° C. or higher, as the lower limit. The humidity varies depending on the temperature, but the lower limit is 0% or more, preferably 25% or more, particularly preferably 50% or more, and the upper limit is 100% or less.

  When the hollow fiber membrane is produced by the wet film-forming method according to the present invention, the winding speed is a factor of production conditions, the shape of the nozzle, the composition of the spinning stock solution, the composition of the internal and external coagulating solutions, the stock solution and Although the speed may vary depending on the temperature of each coagulation liquid, a speed of approximately 600 m / hour to 9,000 m / hour is selected.

By using the wet film formation method according to the present invention, an aromatic ether polymer film having sufficient strength and elongation to be used as a film can be obtained. In addition, by utilizing concentration-induced phase separation in the wet film-forming method of the present invention, it is possible to easily manufacture a porous film structure having an inclined structure, which is difficult to obtain by a melt film-forming method using temperature-induced phase separation. Thus, it is possible to impart high water permeability to the obtained membrane of the present invention.
In the wet film-forming method according to the present invention, after the solidification by the coagulating liquid, the solvent removal can be promoted by immersing in a solvent removal tank in order to increase the strength of the film. The solvent removal liquid is a solvent that can remove the remaining solvent after concentration-induced phase separation by the coagulation liquid, and any solvent that does not dissolve the membrane can be used. In general, water, ethanol or the like is often used.

The film forming temperature in the wet film forming method according to the present invention is a temperature at which the membrane stock solution and the coagulating liquid are brought into contact with each other to cause concentration-induced phase separation. That is, it depends on the temperature of the membrane stock solution, the temperature of the coagulation solution, the temperature of the double spinning nozzle if it is a hollow fiber membrane, and the temperature of a metal plate that supports membrane formation if it is a flat membrane. The lower limit of the film forming temperature is 20 ° C. or higher, preferably 25 ° C. or higher, particularly preferably 30 ° C. or higher. The upper limit is a temperature below each boiling point of the membrane stock solution or coagulation liquid, preferably a temperature lower by 5 ° C. or more from each boiling point, particularly preferably a temperature lower by 10 ° C. or more from the boiling point. In particular, when the film stock solution and the coagulating liquid are in contact with each other within a film forming temperature of 80 ° C. or higher and a temperature lower than each boiling point by 10 ° C. or higher, a particularly high strength film can be obtained.
The undried aromatic ether polymer film obtained by the wet film formation method has a temperature at which film breakage during drying does not occur, for example, within a temperature range of 20 ° C. or more to the melting temperature of the aromatic ether polymer or less. Dry with. The drying temperature is preferably 50 ° C. or higher and 150 ° C. or lower, more preferably 60 ° C. or higher and 140 ° C. or lower, and particularly preferably 70 ° C. or higher and 130 ° C. or lower. The time required for drying is determined by the relationship with the drying temperature, but is generally selected from 0.01 hours to 48 hours.

（式中、Ｒ^１７およびＲ^１８は炭素数１〜１４のアルキル基またはアラルキル基を表す。アルキル基の水素原子またはアラルキル基の水素原子は炭素数１〜１０のアルコキシ基によって置換されていてもよい。式中ｖおよびｗは繰り返し単位を表す。）
その中でも、ポリ（メタ）アクリル酸やポリ（メタ）アクリル酸エステルなど用いることができる。好ましくはポリアクリル酸、ポリメタクリル酸、ポリアクリル酸メチル、ポリアクリル酸エチル、ポリメタアクリル酸メチル、ポリメタクリル酸エチルおよびこれら２つ以上組み合わせた共重合体が良い。必要に応じて高分子末端および／または主鎖中にエステル化、エーテル化、エポキシ化など各種変性を実施することができる。また、免疫グロブリンの静電気的な特性との相性から、アミノ基、モノアルキルアミノ基、ジアルキルアミノ基、カルボキシル基、スルフォニル基、スルホン酸基などの化学構造を必要に応じて導入しても良い。
本発明に係わる（メタ）アクリル系高分子膜は、主としてポリ（メタ）アクリル酸エステルからなるものであるが、ポリ（メタ）アクリル酸エステルの特性を損なわない範囲で他の高分子量物質や添加物を含有していてもよい。これらの高分子を二種以上、組み合わせて実施することも可能である。 The (meth) acrylic polymer film according to the present invention is not particularly limited, and examples of the (meth) acrylic polymer include those represented by the following formula (17).
[Chemical 17]

(In the formula, R ¹⁷ and R ¹⁸ represent an alkyl group or aralkyl group having 1 to 14 carbon atoms. The hydrogen atom of the alkyl group or the hydrogen atom of the aralkyl group may be substituted by an alkoxy group having 1 to 10 carbon atoms. (Wherein v and w represent repeating units)
Among them, poly (meth) acrylic acid and poly (meth) acrylic acid ester can be used. Preferred are polyacrylic acid, polymethacrylic acid, polymethyl acrylate, polyethyl acrylate, polymethyl methacrylate, polyethyl methacrylate, and a copolymer of two or more thereof. Various modifications such as esterification, etherification, and epoxidation can be performed on the polymer terminal and / or main chain as necessary. Moreover, chemical structures such as an amino group, a monoalkylamino group, a dialkylamino group, a carboxyl group, a sulfonyl group, and a sulfonic acid group may be introduced as necessary in view of compatibility with the electrostatic properties of immunoglobulins.
The (meth) acrylic polymer film according to the present invention is mainly composed of poly (meth) acrylic acid ester, but other high molecular weight substances and additives are added within the range that does not impair the properties of poly (meth) acrylic acid ester. You may contain the thing. It is also possible to carry out by combining two or more of these polymers.

  The weight average molecular weight of the (meth) acrylic polymer according to the present invention is 5,000 or more, preferably 10,000 or more, particularly preferably 20,000 or more as the lower limit, and 1,000,000 or less, preferably 500,000 as the upper limit. Hereinafter, 300,000 or less is particularly preferable. Within this range, sufficient strength and film formability can be obtained.

  In the present invention, it is preferable to use a hydrophilic polymer together with the (meth) acrylic polymer in order to control the pore size of the membrane and to impart hydrophilicity. Examples of the hydrophilic polymer include polyvinyl pyrrolidone, polyethylene glycol, polypropylene glycol, polyethylene oxide, polyethylene glycol-polypropylene glycol block copolymer, polyvinyl alcohol, poly (meth) acrylamide, poly-N, N-dimethyl (meth) acrylamide, Examples thereof include poly-N-isopropyl (meth) acrylamide, polyhydroxy acrylate, polyhydroxy methacrylate, carboxymethyl cellulose, starch, corn starch, polychitosan, and polychitin. Among them, polyvinyl pyrrolidone is particularly preferable for improving the hydrophilicity of the entire film because of its good compatibility with (meth) acrylic polymers.

  The weight average molecular weight of the hydrophilic polymer imparting hydrophilicity to the (meth) acrylic polymer film according to the present invention is 1,000 or more, preferably 5,000 or more as the lower limit, and 2 million or less as the upper limit. Preferably it is 1 million or less, and particularly preferably 500,000 or less. For example, various grades of polyvinyl pyrrolidone are commercially available from BASF and have a weight average molecular weight of 9,000 (K17), and similarly 45,000 (K30), 450,000 (K60), 900,000. (K80) and 1,200,000 (K90) are preferably used, and in order to obtain the intended use, characteristics, and structure, each may be used alone or in combination of two or more. .

The molecular weight cutoff of the (meth) acrylic polymer membrane according to the present invention is only required to sufficiently separate the immunoglobulin monomer and the dimer, and the lower limit is 100,000 or more, preferably 150,000 or more, more preferably 250,000 or more is good, and the upper limit should be set to less than 500,000, preferably 450,000 or less, more preferably 400,000 or less. If the molecular weight cut-off is less than 100,000, there is a problem that the amount of immunoglobulin permeated decreases, and if it is more than 500,000, both the immunoglobulin monomer and dimer permeate through the membrane. Decreases.
The molecular weight cut-off according to the present invention includes proteins such as albumin (66,000), γ-globulin (160,000), catalase (232,000), ferritin (440,000), thyroglobulin (669,000), and PEG. Then, dead end filtration is performed using dextran or the like, and the molecular weight is calculated as the molecular weight at which the rejection is 90% from the relationship between the molecular weight and the rejection.

  The method for producing the (meth) acrylic polymer film according to the present invention is not limited in any way, and examples thereof include a wet film forming method. The wet film formation method is a method in which a membrane stock solution obtained by dissolving a membrane material in a good solvent is brought into contact with a coagulation liquid composed of another solvent that is miscible with the good solvent in the membrane stock solution but is not compatible with the membrane material. In this method, concentration-induced phase separation is generated from the contact surface to obtain a membrane.

  The good solvent used in the wet film forming method for obtaining the (meth) acrylic polymer film according to the present invention is not limited as long as it is miscible with water, but 10 g or more is dissolved in 100 g of pure water at 20 ° C. Preferably, it is possible to dissolve 5% by weight or more of a (meth) acrylic polymer as a film material, and more preferably, specifically, N-methyl-2-pyrrolidone, N, N-dimethylacetamide, dimethyl Examples thereof include sulfoxide, N, N-dimethylformamide, acetone, and γ-butyrolactone. From the viewpoint of danger, safety and toxicity, dimethyl sulfoxide, N, N-dimethylacetamide, and N-methyl-2-pyrrolidone are preferably used. These solvents can be used alone or in combination of two or more.

The film stock solution used in the wet film forming method for obtaining the (meth) acrylic polymer film according to the present invention is not limited as long as a (meth) acrylic polymer film having the target structure and performance can be produced. Regarding the concentration of the (meth) acrylic polymer in the membrane stock solution, the film formability improves as the concentration is increased, but conversely, the porosity of the membrane decreases and the water permeability tends to decrease. Therefore, when the whole membrane stock solution is 100% by weight, the concentration range of the (meth) acrylic polymer varies depending on the molecular weight, but the lower limit is 2% by weight or more, preferably 5% by weight or more, particularly preferably 10% by weight. That's it. Further, an upper limit of 50% by weight or less, preferably 40% by weight or less, particularly preferably 30% by weight or less, is preferably used.
In addition, the lower limit of the temperature of the membrane stock solution is preferably 0 ° C. or higher, preferably 10 ° C. or higher, particularly preferably 25 ° C. or higher, and the upper limit is a good solvent boiling point or lower in the membrane stock solution. If it is this temperature condition, the viscosity suitable for processing into a film | membrane preferable as a film | membrane stock solution can be obtained.
Addition of antioxidants, crystal nucleating agents, antistatic agents, flame retardants, lubricants, ultraviolet absorbers, etc., depending on the purpose, as long as the membrane undiluted solution used in the present invention does not affect the performance of the membrane to be produced. Mixing agents may be used.

When a hydrophilic polymer is used in the wet film formation method for obtaining the (meth) acrylic polymer film according to the present invention, its role is mainly to promote and form the porous structure of the outer porous support layer portion. There is a thickening effect of the membrane stock solution. The amount of the hydrophilic polymer added to the membrane stock solution can be adjusted as appropriate in order to achieve stable film formation. When the viscosity of the film stock solution is low, film breakage or film breakage may occur at the time of film formation, and the film formability may become unstable. Conversely, if the viscosity of the membrane stock solution is too high, the porous support layer cannot be sufficiently grown, the porosity of the porous structure of the outer layer will be insufficient, and it will be difficult to obtain a membrane with the desired high permeability. . Furthermore, there is a concern that the stock solution discharged from the die may cause a melt fracture due to an increase in the viscosity of the membrane stock solution.
In the wet film formation method for obtaining the (meth) acrylic polymer film according to the present invention, the upper limit of the concentration of the hydrophilic polymer in the film stock solution is an optimum value according to the type and molecular weight of the hydrophilic polymer to be used. Is usually 40% by weight or less, preferably 30% by weight or less.

  As a coagulating liquid used in the wet film forming method for obtaining the (meth) acrylic polymer film according to the present invention, a substance capable of forming a film from the contact surface by causing concentration-induced phase separation when in contact with the film stock solution. For example, pure water, a monoalcohol solvent, a polyol solvent, or a mixture of two or more of these is preferably used. Examples of the monoalcohol solvent include methanol, ethanol, propanol and the like. Examples of the polyol solvent include ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, glycerin, propylene glycol, 1,2-butanediol, 1,4-butanediol, and the like. In the coagulation liquid, polyvinyl alcohol, polyethylene glycol, polypropylene glycol, polyethylene oxide, polyethylene glycol-polypropylene glycol block copolymer, poly (meth) acrylamide, polyvinylpyrrolidone, polyhydroxyacrylate, polyhydroxymethacrylate, poly (meth) acrylic acid, Polymethacrylic acid, polyitaconic acid, polyfumaric acid, polycitraconic acid, poly-p-styrene sulfonic acid, sodium poly-p-styrene sulfonate, N, N-dimethyl (meth) acrylamide, carboxymethylcellulose, starch, corn starch, polychitosan It is also possible to add a water-soluble polymer such as polychitin. Although it depends on the molecular weight and the amount of the water-soluble polymer to be added, the filtration performance can be improved by adding these. The coagulation liquid may contain a good solvent such as N-methyl-2-pyrrolidone, N, N-dimethylacetamide, dimethyl sulfoxide, N, N-dimethylformamide, and γ-butyrolactone. In particular, when using a coagulation liquid containing a good solvent in a non-solvent, the composition differs depending on the composition of the membrane stock solution, the contact temperature between the membrane stock solution and the coagulation liquid, etc. In this case, 90% by weight or less is preferable as the weight% of the good solvent. Within this range, concentration-induced phase separation necessary and sufficient for forming a film can be sufficiently achieved.

  In the wet film forming method for obtaining the (meth) acrylic polymer film according to the present invention, the coagulating liquid (hereinafter referred to as the internal coagulating liquid) that passes through the inside of the yarn during the production of the hollow fiber membrane is the same solution as the above external coagulating liquid. Alternatively, it may be produced by a dry / wet film forming method in which a gas such as air, nitrogen or ammonia gas is introduced.

  The film forming temperature in the wet film forming method for obtaining the (meth) acrylic polymer film according to the present invention is not limited as long as it is a temperature at which the film stock solution and the coagulating liquid are brought into contact with each other to cause concentration induced phase separation. However, the lower limit of the film formation temperature is 0 ° C. or higher, preferably 10 ° C. or higher, particularly preferably 25 ° C. or higher. The upper limit is a temperature below each boiling point of the membrane stock solution or coagulation liquid, preferably a temperature lower by 5 ° C. or more from each boiling point, particularly preferably a temperature lower by 10 ° C. or more from the boiling point. For hollow fiber membranes, it depends on the temperature of the double nozzle. In a flat membrane, it is determined by the coagulation liquid temperature.

  The membrane stock solution and coagulation solution used in the wet film formation method for obtaining the (meth) acrylic polymer membrane according to the present invention, particularly the coagulation solution (hereinafter referred to as internal coagulation solution) that passes through the inside of the yarn during the production of the hollow fiber membrane, is uniformly dissolved. It is desirable to remove the dissolved gas later. By removing the dissolved gas, film defects due to foaming of the dissolved gas can be remarkably improved. Further, by removing oxygen from the dissolved gas, the oxidation reaction to the material due to film processing at a high temperature is reduced. This step may be omitted when no gas is dissolved in the membrane stock solution, the coagulation solution, and the internal coagulation solution. In addition, when a gas such as air, nitrogen, ammonia gas or the like is used as a coagulant as a dry / wet film forming method, this step is not performed.

When producing a hollow fiber membrane by a wet film formation method for obtaining a (meth) acrylic polymer membrane according to the present invention, in order to further accelerate the coagulation by the membrane stock solution and the internal coagulation solution from the double spinning nozzle, A tank (hereinafter referred to as a coagulation tank) is provided directly below, and can be brought into contact with a coagulation liquid (hereinafter referred to as an external coagulation liquid) filled in the coagulation tank.
When producing a hollow fiber membrane by a wet film forming method for obtaining a (meth) acrylic polymer membrane according to the present invention, the hollow fiber membrane can be freely structured not only in a uniform structure but also in various non-uniform structures. In order to control, the distance from the spinning nozzle to the external coagulation liquid surface (hereinafter referred to as idle running distance) and the temperature and humidity of the space from the spinning nozzle to the external coagulation liquid surface can be adjusted. Although there is no limitation as long as the temperature and humidity of the space can be adjusted, for example, the lower limit of the free running distance is 0.001 m or more, preferably 0.005 m or more, particularly preferably 0.01 m or more, and the upper limit is 2.0 m or less, preferably Is 1.5 m or less, particularly preferably 1.2 m or less. The temperature in the space from the spinning nozzle to the external solidification surface is 10 ° C. or higher, preferably 20 ° C. or higher, particularly preferably 25 ° C. or higher as the lower limit. The humidity varies depending on the temperature, but the lower limit is 0% or more, preferably 10% or more, particularly preferably 30% or more, and the upper limit is 100% or less.

  The winding speed in the case of producing a hollow fiber membrane by a wet film forming method for obtaining the (meth) acrylic polymer membrane according to the present invention is the production conditions, factors such as the spinneret, the composition of the spinning dope, the internal Although the speed may vary depending on the composition of the coagulation liquid and the external coagulation liquid, the temperature of the raw liquid and each coagulation liquid, a speed of approximately 300 m / hour to 9,000 m / hour is selected.

  In the wet film forming method for obtaining the (meth) acrylic polymer film according to the present invention, the solvent removal can be promoted by solidifying with a coagulating liquid and then immersing in a solvent removing tank to increase the strength of the film. The solvent removal liquid is a solvent that can remove the remaining solvent after concentration-induced phase separation by the coagulation liquid, and any solvent that does not dissolve the membrane can be used. In general, water, ethanol or the like is often used.

  The drying temperature of the undried (meth) acrylic polymer film of the present invention obtained by the wet film formation method according to the present invention is not limited as long as it does not cause film breakage during drying. The drying is performed within a temperature range from 20 ° C. or more to the melting temperature of the (meth) acrylic polymer. The preferable drying temperature is 30 ° C. or higher as the lower limit, preferably 40 ° C. or higher, and 80 ° C. or lower, preferably 70 ° C. or lower as the upper limit. The time required for drying is determined by the relationship with the drying temperature, but is generally selected from 0.01 hours to 48 hours.

（式中、Ｒ^１９およびＲ^２０は、水素またはメチル基を表す。Ｒ^２１は炭素数１〜１４のアルキル基またはアラルキル基を表す。アルキル基の水素原子またはアラルキル基の水素原子は、炭素数１〜１０のアルコキシ基によって置換されていてもよい。式中ｘおよｙは繰り返し単位を表す。）
その中でも、ポリ（メタ）アクリロニトリルやポリ（メタ）アクリロニトリル酸エステルなど用いることができる。好ましくはポリアクリロニトリル、ポリメタアクリロニトリルが良い。 The (meth) acrylonitrile film according to the present invention is not particularly limited, and examples of the (meth) acrylonitrile polymer include those represented by the following formula (18).
[Chemical 18]

(In the formula, R ¹⁹ and R ²⁰ represent hydrogen or a methyl group. R ²¹ represents an alkyl group or an aralkyl group having 1 to 14 carbon atoms. The hydrogen atom of the alkyl group or the hydrogen atom of the aralkyl group represents the number of carbon atoms. (It may be substituted by an alkoxy group of 1 to 10. In the formula, x and y represent a repeating unit.)
Among these, poly (meth) acrylonitrile, poly (meth) acrylonitrile acid ester, and the like can be used. Polyacrylonitrile and polymethacrylonitrile are preferable.

  The monomer composition constituting the (meth) acrylonitrile polymer according to the present invention has a (meth) acrylonitrile content of at least 50% by weight, preferably 60% by weight or more, and is copolymerizable with (meth) acrylonitrile. The content of one or two or more kinds of vinyl compounds is 50% by weight or less, preferably 40% by weight or less. The vinyl compound is not particularly limited as long as it is a known compound having copolymerizability with (meth) acrylonitrile, and preferred copolymer components include (meth) acrylic acid and methyl (meth) acrylate. , Ethyl (meth) acrylate, itaconic acid, vinyl acetate, sodium (meth) acrylsulfonate, sodium p (para) -styrenesulfonate, hydroxyethyl methacrylate, ethyl triethylammonium methacrylate, ethyl trimethylammonium methacrylate Examples include chloride and vinyl pyrrolidone. Examples thereof include acrylonitrile-methyl acrylate-PVP copolymer.

Various modifications such as esterification, etherification, and epoxidation can be performed on the polymer terminal and / or main chain as necessary. Moreover, chemical structures such as an amino group, a monoalkylamino group, a dialkylamino group, a carboxyl group, a sulfonyl group, and a sulfonic acid group may be introduced as necessary in view of compatibility with the electrostatic properties of immunoglobulins.
The (meth) acrylonitrile-based polymer film according to the present invention is mainly composed of a (meth) acrylonitrile-based polymer, but other high molecular weight substances and additives are added within the range not impairing the properties of the (meth) acrylonitrile-based polymer. You may contain the thing. It is also possible to carry out by combining two or more of these polymers.

  The lower limit of the weight average molecular weight of the (meth) acrylonitrile polymer according to the present invention is 5,000 or more, preferably 10,000 or more, particularly preferably 20,000 or more, and the upper limit is 1,000,000 or less, preferably 500,000. Hereinafter, 300,000 or less is particularly preferable. Within this range, sufficient strength and film formability can be obtained.

  In the present invention, it is preferable to use a hydrophilic polymer together with the (meth) acrylonitrile polymer in order to control the pore size of the membrane and to impart hydrophilicity. Examples of the hydrophilic polymer include polyvinyl pyrrolidone, polyethylene glycol, polypropylene glycol, polyethylene oxide, polyethylene glycol-polypropylene glycol block copolymer, polyvinyl alcohol, poly (meth) acrylamide, poly-N, N-dimethyl (meth) acrylamide, Examples thereof include poly-N-isopropyl (meth) acrylamide, polyhydroxy acrylate, polyhydroxy methacrylate, carboxymethyl cellulose, starch, corn starch, polychitosan, and polychitin. Among them, polyvinyl pyrrolidone has a good compatibility with the (meth) acrylonitrile-based polymer, and is particularly preferable for improving the hydrophilicity of the entire film.

  The weight average molecular weight of the hydrophilic polymer that imparts hydrophilicity to the (meth) acrylonitrile-based polymer film according to the present invention is 1,000 or more, preferably 5,000 or more as a lower limit, and 2 million as an upper limit. Below, it is preferably 1,000,000 or less, particularly preferably 500,000 or less. For example, various grades of polyvinyl pyrrolidone are commercially available from BASF and have a weight average molecular weight of 9,000 (K17), and similarly 45,000 (K30), 450,000 (K60), 900,000. (K80) and 1,200,000 (K90) are preferably used, and in order to obtain the intended use, characteristics, and structure, each may be used alone or in combination of two or more. .

The molecular weight cutoff of the (meth) acrylonitrile-based polymer membrane according to the present invention is only required to sufficiently separate the immunoglobulin monomer and dimer, and the lower limit is 100,000 or more, preferably 150,000 or more, more preferably 25. It is necessary to set the upper limit to less than 500,000, preferably 450,000 or less, and more preferably 400,000 or less. If the molecular weight cut-off is less than 100,000, there is a problem that the amount of immunoglobulin permeated decreases, and if it is more than 500,000, both the immunoglobulin monomer and dimer permeate through the membrane. Decreases.
The molecular weight cut-off according to the present invention includes proteins such as albumin (66,000), γ-globulin (160,000), catalase (232,000), ferritin (440,000), thyroglobulin (669,000), and PEG. Then, dead end filtration is performed using dextran or the like, and the molecular weight is calculated as the molecular weight at which the rejection is 90% from the relationship between the molecular weight and the rejection.

The method for producing the (meth) acrylonitrile polymer film according to the present invention is not limited in any way, and examples thereof include a wet film forming method. The wet film formation method is to bring a membrane stock solution in which a membrane material is dissolved in a good solvent into contact with a coagulation liquid composed of another solvent that is miscible with the good solvent in the membrane stock solution but is incompatible with the membrane material. In this method, concentration-induced phase separation is generated from the contact surface to obtain a membrane.
The good solvent used in the wet film formation method for obtaining the (meth) acrylonitrile polymer film according to the present invention is not limited as long as it is miscible with water, but 10 g or more is dissolved in 100 g of pure water at 20 ° C. It is possible to use a film material that dissolves 5% by weight or more of the (meth) acrylonitrile polymer as a film material. For example, N-methyl-2-pyrrolidone, N, N-dimethylacetamide, dimethyl sulfoxide, N, N-dimethylformamide, acetone, γ-butyrolactone and the like can be mentioned. From the viewpoint of danger, safety and toxicity, dimethyl sulfoxide, N, N-dimethylacetamide, and N-methyl-2-pyrrolidone are preferably used. These solvents can be used alone or in combination of two or more.

The membrane stock solution used in the wet film formation method for obtaining the (meth) acrylonitrile polymer film according to the present invention is not limited as long as it can produce a (meth) acrylonitrile polymer having the target structure and performance, but the concentration is not limited. As the film is raised, the film formability improves, but conversely, the porosity of the film decreases and the water permeability tends to decrease. Therefore, when the whole membrane stock solution is 100% by weight, the concentration range of the (meth) acrylonitrile polymer varies depending on the molecular weight, but the lower limit is 2% by weight or more, preferably 5% by weight or more, particularly preferably 10% by weight. % Or more. Further, an upper limit of 50% by weight or less, preferably 40% by weight or less, particularly preferably 30% by weight or less, is preferably used. When the concentration of the (meth) acrylonitrile polymer is less than 2% by weight, the viscosity of the stock solution is low and tends to be difficult to form. When it is higher than 50% by weight, the viscosity of the stock solution is too high and film formation is difficult. It tends to be. It is also possible to add multiple non-solvents such as water, salts, alcohols, ethers, ketones, glycols, etc. for the purpose of controlling the viscosity of the stock solution and the dissolution state, and the type and amount of addition can be changed depending on the combination. Just decide.
In addition, the lower limit of the temperature of the membrane stock solution is preferably 0 ° C. or higher, preferably 10 ° C. or higher, particularly preferably 25 ° C. or higher, and the upper limit is a good solvent boiling point or lower in the membrane stock solution. If it is this temperature condition, the viscosity suitable for processing into a film | membrane preferable as a film | membrane stock solution can be obtained.
Addition of antioxidants, crystal nucleating agents, antistatic agents, flame retardants, lubricants, ultraviolet absorbers, etc., depending on the purpose, as long as the membrane undiluted solution used in the present invention does not affect the performance of the membrane to be produced. Mixing agents may be used.

When a hydrophilic polymer is used in a wet film forming method for obtaining a (meth) acrylonitrile-based polymer film according to the present invention, its role is mainly to promote the porous structure of the outer porous support layer part. There is a thickening effect of the membrane stock solution. The amount of the hydrophilic polymer added to the membrane stock solution can be adjusted as appropriate in order to achieve stable film formation. When the viscosity of the membrane stock solution is low, yarn breakage and yarn shaking may occur during film formation, which may make the yarn production unstable. Conversely, if the viscosity of the membrane stock solution is too high, the porous support layer cannot be sufficiently grown, the porosity of the porous structure of the outer layer will be insufficient, and it will be difficult to obtain a membrane with the desired high permeability. . Furthermore, there is a concern that the stock solution discharged from the die may cause a melt fracture due to an increase in the viscosity of the membrane stock solution.
In the wet film-forming method for obtaining the (meth) acrylonitrile polymer film according to the present invention, the upper limit of the concentration of the hydrophilic polymer in the membrane stock solution is an optimum value according to the type and molecular weight of the hydrophilic polymer to be used. Is usually 40% by weight or less, preferably 30% by weight or less.

  The coagulating liquid used in the wet film-forming method for obtaining the (meth) acrylonitrile-based polymer film according to the present invention is a substance capable of causing concentration-induced phase separation when in contact with the film stock solution and forming a film from the contact surface. For example, pure water, a monoalcohol solvent, a polyol solvent, or a mixture of two or more of these is preferably used. Examples of the monoalcohol solvent include methanol, ethanol, propanol and the like. Examples of the polyol solvent include ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, glycerin, propylene glycol, 1,2-butanediol, 1,4-butanediol, and the like. In the coagulation liquid, polyvinyl alcohol, polyethylene glycol, polypropylene glycol, polyethylene oxide, polyethylene glycol-polypropylene glycol block copolymer, poly (meth) acrylamide, polyvinylpyrrolidone, polyhydroxyacrylate, polyhydroxymethacrylate, poly (meth) acrylic acid, Polymethacrylic acid, polyitaconic acid, polyfumaric acid, polycitraconic acid, poly-p-styrene sulfonic acid, sodium poly-p-styrene sulfonate, N, N-dimethyl (meth) acrylamide, carboxymethylcellulose, starch, corn starch, polychitosan It is also possible to add a water-soluble polymer such as polychitin. Further, a liquid that does not dissolve the polymer such as aliphatic hydrocarbons such as n-hexane and n-heptane may be used. Although it depends on the molecular weight and the amount of the water-soluble polymer to be added, the filtration performance can be improved by adding these. In addition, a good solvent such as N-methyl-2-pyrrolidone, N, N-dimethylacetamide, dimethyl sulfoxide, N, N-dimethylformamide, γ-butyrolactone, propylene carbonate, ethylene carbonate may be contained in the coagulation liquid. Is possible. In particular, when using a coagulation liquid containing a good solvent in a non-solvent, the composition differs depending on the composition of the membrane stock solution, the contact temperature between the membrane stock solution and the coagulation liquid, etc. In this case, 90% by weight or less is preferable as the weight% of the good solvent. Within this range, concentration-induced phase separation necessary and sufficient for forming a film can be sufficiently achieved.

  The film formation temperature in the wet film formation method for obtaining the (meth) acrylonitrile polymer film according to the present invention is not limited as long as it is a temperature at which the membrane stock solution and the coagulation liquid are brought into contact with each other to cause concentration induced phase separation. However, the lower limit of the film formation temperature is 0 ° C. or higher, preferably 10 ° C. or higher, particularly preferably 25 ° C. or higher. The upper limit is a temperature below each boiling point of the membrane stock solution or coagulation liquid, preferably a temperature lower by 5 ° C. or more from each boiling point, particularly preferably a temperature lower by 10 ° C. or more from the boiling point. For hollow fiber membranes, it depends on the temperature of the double nozzle. In a flat membrane, it is determined by the coagulation liquid temperature.

  The membrane stock solution used in the wet film-forming method for obtaining the (meth) acrylonitrile polymer membrane according to the present invention, the coagulating solution, especially the coagulating solution passing through the inside of the yarn during the production of the hollow fiber membrane (hereinafter referred to as the internal coagulating solution) is uniformly dissolved. It is desirable to remove the dissolved gas later. By removing the dissolved gas, film defects due to foaming of the dissolved gas can be remarkably improved. Further, by removing oxygen from the dissolved gas, the oxidation reaction to the material due to film processing at a high temperature is reduced. This step may be omitted when no gas is dissolved in the membrane stock solution, the coagulation solution, and the internal coagulation solution.

In the case of producing a hollow fiber membrane by a wet film forming method for obtaining a (meth) acrylonitrile-based polymer membrane according to the present invention, in order to further promote the coagulation by the membrane stock solution and the internal coagulation solution discharged from the double spinning nozzle, A tank (hereinafter referred to as a coagulation tank) is provided directly below, and can be brought into contact with a coagulation liquid (hereinafter referred to as an external coagulation liquid) filled in the coagulation tank.
When producing a hollow fiber membrane by a wet film forming method for obtaining a (meth) acrylonitrile-based polymer membrane according to the present invention, the cross-sectional structure of the hollow fiber membrane can be freely structured not only to a uniform structure but also to various non-uniform structures. In order to control, the distance from the spinning nozzle to the external coagulation liquid surface (hereinafter referred to as idle running distance) and the temperature and humidity of the space from the spinning nozzle to the external coagulation liquid surface can be adjusted. Although there is no limitation as long as the temperature and humidity of the space can be adjusted, for example, the lower limit of the free running distance is 0.001 m or more, preferably 0.005 m or more, particularly preferably 0.01 m or more, and the upper limit is 2.0 m or less, preferably Is 1.5 m or less, particularly preferably 1.2 m or less. The temperature in the space from the spinning nozzle to the external solidification surface is 10 ° C. or higher, preferably 20 ° C. or higher, particularly preferably 25 ° C. or higher as the lower limit. The humidity varies depending on the temperature, but the lower limit is 0% or more, preferably 10% or more, particularly preferably 30% or more, and the upper limit is 100% or less.

  The winding speed in the case of producing a hollow fiber membrane by a wet film forming method for obtaining the (meth) acrylonitrile-based polymer membrane according to the present invention is various factors that are production conditions, the shape of the spinneret, the composition of the spinning dope, the internal Although the speed may vary depending on the composition of the coagulation liquid and the external coagulation liquid, the temperature of the raw liquid and each coagulation liquid, a speed of approximately 300 m / hour to 9,000 m / hour is selected.

In the wet film forming method for obtaining the (meth) acrylonitrile-based polymer film according to the present invention, after coagulation with the coagulating liquid, the solvent removal can be promoted by immersing in a solvent removal tank in order to increase the strength of the film. The solvent removal liquid is a solvent that can remove the remaining solvent after concentration-induced phase separation by the coagulation liquid, and any solvent that does not dissolve the membrane can be used. In general, water, ethanol or the like is often used.
The drying temperature of the undried (meth) acrylonitrile-based polymer film of the present invention obtained by the wet film forming method according to the present invention is not limited as long as it does not cause film breakage during drying. The drying is performed within a temperature range from 20 ° C. or more to the melting temperature of the (meth) acrylonitrile polymer. A preferable drying temperature is 30 ° C. or higher and 75 ° C. or lower. The time required for drying is determined by the relationship with the drying temperature, but is generally selected from 0.01 hours to 48 hours.

The fluorine-based polymer film according to the present invention is mainly composed of a fluorine-based polymer, but may contain other high molecular weight substances and additives as long as the characteristics of the fluorine-based polymer are not impaired. It is also possible to carry out by combining two or more of these polymers.
The fluorine-based polymer according to the present invention is a homopolymer of vinylidene fluoride, one kind selected from a monomer group of hexafluoropropylene, pentafluoropropylene, tetrafluoroethylene, chlorotrifluoroethylene, and perfluoromethyl vinyl ether, or It is a copolymer of two types of monomers and vinylidene fluoride. Moreover, the said homopolymer and the said copolymer can also be mixed and used. Among these, vinylidene fluoride is preferable.

  The weight average molecular weight of the fluoropolymer according to the present invention is preferably 50,000 or more, preferably 100,000 or more, particularly preferably 150,000 or more as a lower limit, and 5 million or less, preferably 2 million or less, particularly preferably as an upper limit. Is preferably 1 million or less. In general, a resin having an average molecular weight exceeding 1,000,000 is difficult to measure by gel permeation chromatography (GPC). Therefore, a viscosity average molecular weight by a viscosity method can be applied as a substitute. If the average molecular weight is less than 50,000, the melt tension at the time of melt molding becomes small and the moldability is deteriorated, and the mechanical strength of the film is lowered. An average molecular weight exceeding 5 million is not preferable because uniform melt-kneading becomes difficult.

  In order to prevent clogging due to the adsorption of immunoglobulin, it is necessary to impart hydrophilicity to the membrane. Examples of the hydrophilization treatment method include, for example, a method in which a fluoropolymer film is immersed in a solution containing a surfactant and then dried to leave the surfactant in the fluoropolymer film. The method of grafting hydrophilic acrylic monomer, methacrylic monomer, etc. on the pore surface of the fluoropolymer film by irradiating the above-mentioned radiation or using a peroxide. A method of mixing, a method of immersing the fluoropolymer film in a solution containing a hydrophilic polymer, and then drying to form a hydrophilic polymer film on the pore surface of the fluoropolymer film. Graft polymerization is most preferable in consideration of the persistence of hydrophilization and the possibility of leakage of hydrophilic additives. In particular, the hydrophilic treatment by the radiation graft polymerization method disclosed in JP-A-62-2179540, JP-A-62-258711, and U.S. Pat. It is preferable in that a uniform hydrophilic layer can be formed on the surface of the pores.

  The hydrophilic monomer used in the graft polymerization of the present invention is not particularly limited as long as it is a hydrophilic monomer having a vinyl group. A monomer having one vinyl group is preferable. Furthermore, (meth) acrylic monomers containing a sulfone group, a carboxyl group, an amide group, a neutral hydroxyl group, a sulfonyl group, a sulfonyl group, a sulfonic acid group, etc. can be suitably used, but when filtering a solution containing an immunoglobulin Is particularly preferably a monomer containing a neutral hydroxyl group. The hydrophilic monomer according to the present invention is a monomer that is uniformly dissolved when mixed with 1% by volume of pure water at 25 ° C. under atmospheric pressure. For example, a vinyl monomer having a hydroxyl group such as hydroxypropyl acrylate, or a functional group serving as a precursor thereof, a vinyl monomer having an anion exchange group such as triethylammonium ethyl methacrylate, or a cation exchange group such as sulfopropyl methacrylate. And vinyl monomers having an amide bond such as vinyl pyrrolidone. Among them, a vinyl monomer having one or more hydroxyl groups or a functional group serving as a precursor thereof is preferable because the permeability of the immunoglobulin solution is the highest. Specifically, (meth) acrylic acid and polyhydric alcohol esters such as hydroxypropyl acrylate and 2-hydroxyethyl methacrylate, alcohols having an unsaturated bond such as allyl alcohol, and vinyl acetate and vinyl propionate. Examples include enol esters. In addition, a hydrophilic monomer having one vinyl group and a crosslinking agent having two or more vinyl groups are used in a proportion of 20 mol% or more and 1,000 mol% or less with respect to the hydrophilic monomer. Hydrophilicity is sufficiently achieved by copolymerization by a polymerization method.

  The cross-linking agent used according to the present invention is a cross-linking agent having two or more vinyl groups that can be copolymerized with the hydrophilic monomer, and is introduced by contacting the membrane simultaneously with the hydrophilic monomer. The number average molecular weight of the crosslinking agent is 200 or more, preferably 250 or more, more preferably 300 or more as the lower limit, and the upper limit is 2,000 or less, preferably 1,000 or less, more preferably 600 or less. is there. When the number average molecular weight of the crosslinking agent is 200 or more and 2,000 or less, a high filtration rate of the immunoglobulin solution is obtained, which is preferable. In the present invention, any crosslinking agent having two or more vinyl groups can be used, but a hydrophilic crosslinking agent is preferred. Here, the hydrophilic cross-linking agent is a cross-linking agent that is uniformly dissolved when mixed with 1% by volume of pure water at 25 ° C. under atmospheric pressure.

  Specific examples of the crosslinking agent used in the present invention include divinylbenzene derivatives for aromatic systems, methacrylic acid crosslinking agents such as ethylene glycol dimethacrylate and polyethylene glycol dimethacrylate for aliphatic systems, ethylene glycol diacrylate, Examples include (meth) acrylic acid-based crosslinking agents such as polyethylene glycol diacrylate. A crosslinking agent having three reactive groups such as trimethylolpropane trimethacrylate can also be used. In addition, a mixture of two or more kinds of crosslinking agents can be used. In the present invention, it is most preferable to use polyethylene glycol dimethacrylate, polyethylene glycol diacrylate, or a mixture thereof from the viewpoints of immunoglobulin monomer permeability and immunoglobulin dimer removal performance.

  The graft polymerization method according to the present invention is not particularly limited as long as it is a method of generating radicals. For example, radicals are generated in a fluorine-based polymer film by means such as addition of a radiation initiator, ionizing radiation or chemical reaction. Then, starting from the radical, the monomer is graft-polymerized to the film. In the present invention, any means can be employed for generating radicals in the fluorine-based polymer film, but irradiation with ionizing radiation is preferable in order to generate uniform radicals throughout the film. As types of ionizing radiation, γ rays, electron beams, β rays, neutron rays, and the like can be used, but electron beams or γ rays are most preferable for implementation on an industrial scale. The ionizing radiation is obtained from radioactive isotopes such as cobalt 60, strontium 90, and cesium 137, or by an X-ray imaging apparatus, an electron beam accelerator, an ultraviolet irradiation apparatus, or the like.

  The irradiation dose of ionizing radiation according to the present invention is preferably from 1 kGy to 1,000 kGy. If it is less than 1 kGy, radicals are not generated uniformly, and if it exceeds 1,000 kGy, film strength may be reduced. Graft polymerization is generally divided into two types: a pre-irradiation method in which radicals are generated on a film and then contacted with a reactive compound, and a simultaneous irradiation method in which radicals are generated on the film in a state where the film is in contact with the reactive compound. Is done. In the present invention, any method can be applied, but the pre-irradiation method in which oligomer formation is small is most preferable.

  In the present invention, the contact between the fluorine-based polymer film that has generated radicals, the hydrophilic monomer and the cross-linking agent is achieved in the liquid phase even in the gas phase, but in the present invention, the liquid phase in which the grafting reaction proceeds uniformly. The method of contacting with is a preferred method. In order to further promote the graft reaction more uniformly, it is desirable that the hydrophilic monomer and the crosslinking agent are dissolved in a solvent in advance and then contacted with the fluoropolymer film. The solvent for dissolving the hydrophilic monomer and the crosslinking agent is not particularly limited as long as it can be uniformly dissolved. Examples of such solvents include alcohols such as ethanol, isopropanol, and t-butyl alcohol, ethers such as diethyl ether and tetrahydrofuran, ketones such as acetone and 2-butanone, water, and mixtures thereof. .

The graft polymerization according to the present invention uses a reaction solution of 0.3% by volume to 30% by volume with a concentration of a hydrophilic monomer and a cross-linking agent, and 10 × 10 ^{−5 to} 100 × 100 g per 1 g of the fluoropolymer film. It is desirable to carry out the reaction at a rate of × 10 ⁻⁵ m ³ . If graft polymerization is performed within this range, pores are not filled with the hydrophilic layer, and a film having excellent uniformity can be obtained.
The reaction temperature at the time of graft polymerization according to the present invention is not particularly limited as long as the polymerization reaction occurs, but it is generally carried out at 20 ° C. to 80 ° C.
In the graft polymerization according to the present invention, when the fluoropolymer film and the hydrophilic monomer are brought into contact with each other, the hydrophilic monomer may be in any state of gas, liquid or solution, but forms a uniform hydrophilic layer. For this purpose, a liquid or a solution is preferable, and a solution is particularly preferable.

  The hydrophilic fluorine-based polymer membrane according to the present invention introduces an immunoglobulin monomer permeability and an immunoglobulin dimer by introducing a hydrophilic layer having a strong cross-linking structure into a hydrophobic fluorine-based polymer membrane. The blocking property can be realized at a high level. Therefore, the crosslinking agent with respect to the hydrophilic monomer has a lower limit of 20 mol% or more, preferably 30 mol% or more, and an upper limit of 1,000 mol% or less, preferably 500 mol% or less, more preferably 200 mol%. It is good to use in the ratio below%.

In the present invention, a hydrophilic layer is introduced into a hydrophobic fluorine-based polymer membrane to achieve high immunoglobulin monomer permeability. Therefore, the graft ratio grafted onto the hydrophobic fluorine-based polymer membrane is 3% or more as a lower limit, preferably 4% or more, more preferably 5% or more, and the upper limit is 50% or less, preferably 30% or less. More preferably, 20% or less is good. When the graft ratio is less than 3%, the hydrophilicity of the membrane is insufficient, causing a rapid decrease in the filtration rate accompanying protein adsorption. If it exceeds 50%, relatively small pores are filled with the hydrophilic layer, and a sufficient filtration rate cannot be obtained. The graft ratio referred to here is a value defined by the following formula (19).
Graft rate (%) =
(Membrane weight after grafting−membrane weight before grafting) / membrane weight before grafting × 100 (19)

  The degree of hydrophilicity of the fluoropolymer film according to the present invention can be evaluated by the contact angle. The average value of the advancing contact angle and the receding contact angle at 25 ° C. is preferably 60 degrees or less, more preferably 45 degrees or less, and still more preferably 30 degrees or less. Further, as a simple evaluation method, it may be determined that when the fluorine-based polymer membrane is brought into contact with water, water sufficiently permeates into the pores of the membrane and has sufficient hydrophilicity.

The molecular weight cut off of the hydrophilic fluoropolymer membrane according to the present invention is only required to sufficiently separate immunoglobulin monomers and dimers, and the lower limit is 100,000 or more, preferably 150,000 or more, more preferably 250,000. The upper limit is good, and the upper limit should be set to less than 500,000, preferably 450,000 or less, more preferably 400,000 or less. If the molecular weight cut-off is less than 100,000, there is a problem that the amount of immunoglobulin permeated decreases, and if it is more than 500,000, both the immunoglobulin monomer and dimer permeate through the membrane. Decreases.
The molecular weight cut-off according to the present invention includes proteins such as albumin (66,000), γ-globulin (160,000), catalase (232,000), ferritin (440,000), thyroglobulin (669,000), and PEG. Then, dead end filtration is performed using dextran or the like, and the molecular weight is calculated as the molecular weight at which the rejection is 90% from the relationship between the molecular weight and the rejection.

  The method for producing the fluorine-based polymer film according to the present invention is not limited at all, and examples thereof include a melt film forming method and a wet film forming method. The melt film-forming method is a method in which a film material and a plasticizer are uniformly mixed by heating and then phase separation is generated by cooling, and a film is obtained by extracting the plasticizer from the obtained film film. It is. In addition, the wet film formation method involves contacting a membrane stock solution in which a membrane material is dissolved in a good solvent with a coagulation liquid composed of another solvent that is miscible with the good solvent in the membrane stock solution but is incompatible with the membrane material. In this method, concentration-induced phase separation is generated from the contact surface to obtain a membrane.

A typical melt film forming method for obtaining a fluorine polymer film according to the present invention includes the following steps (a) to (c).
(A) A step of heating a composition containing a fluoropolymer and a plasticizer to a temperature equal to or higher than the crystal melting point of the fluoropolymer and uniformly dissolving the composition, and then discharging the composition from a discharge port to form a film;
(B) A non-volatile liquid having partial solubility in the fluoropolymer while taking the film at a take-off speed such that the draft ratio defined in the following formula (20) is 1 or more and 15 or less. In a state where the temperature is heated to 100 ° C. or higher, and the other membrane surface is cooled:
Draft ratio = (film take-off speed) / (discharge speed at composition discharge port) (20)
(C) removing a substantial part of the plasticizer and the non-volatile liquid;

  The polymer concentration used in the melt film-forming method for obtaining the fluorine-based polymer film according to the present invention is 20% by weight or more, preferably 30% by weight or more, as the lower limit in the composition containing the fluorine-based polymer and the plasticizer. More preferred is 35% by weight, and the upper limit is 90% by weight or less, preferably 80% by weight or less, more preferably 70% by weight or less. When the polymer concentration is less than 20% by weight, the film formability deteriorates, and there are problems such as insufficient mechanical strength. If the polymer concentration exceeds 90% by weight, the pore size of the resulting fluoropolymer membrane will be too small and the porosity will be small, so the filtration rate will be low and it will not be practical.

The plasticizer used in the melt film-forming method for obtaining the fluorine-based polymer film according to the present invention includes a homogeneous solution at a temperature equal to or higher than the crystal melting point of the resin when mixed with the fluorine-based polymer in the composition for producing the fluorine-based polymer film. A non-volatile solvent capable of forming is used. The non-volatile solvent mentioned here has a boiling point of 250 ° C. or higher under atmospheric pressure. The plasticizer may be liquid or solid at a room temperature of 20 ° C. Further, a so-called solid-liquid phase separation type plasticizer that has a heat-induced solid-liquid phase separation point at a temperature of room temperature or higher when a homogeneous solution with a fluorine-based polymer is cooled may be used. Some plasticizers have a heat-induced liquid-liquid phase separation point at a temperature equal to or higher than room temperature when a homogeneous solution with a fluoropolymer is cooled. When an agent is used, the obtained fluoropolymer membrane tends to have a large pore size. The plasticizer used here may be a single item or a mixture of a plurality of substances.
The method of measuring the heat-induced solid-liquid phase separation point uses a sample obtained by pre-melting and kneading a predetermined concentration of a composition containing a fluoropolymer and a plasticizer, and performing thermal analysis such as differential scanning calorimetry (DSC). By measuring the exothermic peak temperature of the resin. Further, the method for measuring the crystallization point of the resin can be obtained by thermal analysis in the same manner using a sample obtained by previously kneading the resin as a sample.

As a plasticizer concerning this invention, the plasticizer currently disclosed by the international publication 01/28667 pamphlet is mentioned. That is, the phase separation point depression constant α of the composition defined by the following formula (21) is 0 ° C. or higher, preferably 1 ° C. or higher, more preferably 5 ° C. or higher as the lower limit, and the upper limit is 40 ° C. or lower. A plasticizer with a temperature of preferably 35 ° C. or lower, more preferably 30 ° C. or lower is preferable. When the phase separation point depression constant exceeds 40 ° C., the uniformity and strength of the pore diameter are lowered, which is not preferable.
α = 100 × (T _c ⁰ −T _c ) ÷ (100−C) (21)
α is the phase separation point drop constant (℃)
T _c ⁰ is the crystallization temperature of the fluoropolymer (℃)
T _c is the thermally induced solid-liquid phase separation point (° C.) of the composition
C is the concentration of fluoropolymer in the composition (% by weight)

  Specifically, phthalic acid esters having an ester chain carbon chain length of 7 or less, adipic acid esters, sebacic acid esters, phosphate esters having an ester chain carbon chain length of 8 or less, citrate esters, etc. Can be preferably used, in particular diheptyl phthalate, dicyclohexyl phthalate, dibutyl phthalate, diethyl phthalate, dimethyl phthalate, dibutyl adipate, dibutyl sebacate, triphenyl phosphate, tricresyl phosphate, diphenyl cresyl phosphate, Tri (2-ethylhexyl) phosphate, tributyl phosphate, tributyl acetylcitrate and the like are particularly preferable.

  In the composition used in the present invention, an antioxidant, a crystal nucleating agent, an antistatic agent, a flame retardant, a lubricant, an ultraviolet absorber and the like are added depending on the purpose as long as the performance of the film to be produced is not affected. Mixing agents may be used.

In the melt film-forming method for obtaining a fluoropolymer film according to the present invention, a first method for uniformly dissolving a composition containing a fluoropolymer and a plasticizer is a continuous resin kneading apparatus such as an extruder. In this method, a plasticizer is introduced at an arbitrary ratio while the resin is heated and melted, and screw kneading is performed to obtain a uniform solution. The form of the resin to be charged may be any of powder, granule, and pellet. Moreover, when making it melt | dissolve uniformly by such a method, it is preferable that the form of a plasticizer is a normal temperature liquid. As an extruder, a single screw type extruder, a biaxial different direction screw type extruder, a biaxial same direction screw type extruder, etc. can be used.
A second method for uniformly dissolving a composition containing a fluoropolymer and a plasticizer is to mix and disperse the fluoropolymer and the plasticizer in advance using a stirring device such as a Henschel mixer. This is a method of obtaining a uniform solution by putting a product into a continuous resin kneader such as an extruder and melt-kneading. The form of the composition to be added may be a slurry when the plasticizer is a liquid at room temperature, and may be a powder or a granule when the plasticizer is a solid at room temperature.
The third method of uniformly dissolving a composition containing a fluoropolymer and a plasticizer is a method using a simple resin kneader such as a Brabender or a mill, or a method of melt kneading in another batch type kneading vessel. is there. According to this method, since it is a batch type process, the productivity is not good, but there is an advantage that it is simple and highly flexible.

In the melt film-forming method for obtaining a fluoropolymer film according to the present invention, a composition containing a fluoropolymer and a plasticizer is heated and dissolved at a temperature equal to or higher than the crystal melting point of the fluoropolymer, After the step (a) of extruding from the discharge port of the circular die or the annular nozzle into a flat membrane shape or a hollow fiber shape, the process proceeds to the step (b) in which it is cooled and solidified to perform the molding. Form a structure.
In the melt film-forming method for obtaining a fluorine-based polymer film according to the present invention, a composition containing a fluorine-based polymer and a plasticizer that are uniformly heated and dissolved is discharged from a discharge port, and a draft defined by the following formula (22) is used. While taking the film at a take-off speed such that the ratio is 1 or more and 15 or less, a non-volatile liquid having partial solubility is brought into contact with the fluoropolymer to form a film.
Draft ratio = (film take-off speed) / (discharge speed at composition outlet) (22)

The draft ratio is preferably 1.5 or more, more preferably 2 or more as the lower limit, and preferably 10 or less, more preferably 7 or less as the upper limit. When the draft ratio is less than 1, no tension is applied to the film and the moldability is lowered. When the draft ratio exceeds 15, the film is stretched and it is difficult to form a sufficiently thick coarse structure layer. The discharge speed at the discharge port of the composition said here is given by the following formula (23).
Discharge speed at the discharge port of the composition =
(Volume of composition discharged per unit time) / (area of discharge port) (23)

The lower limit of the discharge speed is 1 m / min or more, preferably 3 m / min or more, and the upper limit is 60 m / min or less, more preferably 40 m / min or less. When the discharge speed is less than 1 m / min, in addition to a decrease in productivity, problems such as a large fluctuation in discharge amount occur. On the contrary, when the discharge speed exceeds 60 m / min, since the discharge amount is large, turbulent flow may occur at the discharge port, and the discharge state may become unstable. The take-up speed can be set in accordance with the discharge speed. The lower limit is 1 m / min or more, preferably 3 m / min or more, and the upper limit is 200 m / min or less, preferably 150 m / min or less. . When the take-up speed is less than 1 m / min, productivity and moldability are deteriorated, and when the take-up speed exceeds 200 m / min, the cooling time is shortened, and the tension on the film is increased, so that the film is broken. It becomes easy to get up.

  In the present invention, an extraction solvent is used to remove the plasticizer. The extraction solvent is preferably a poor solvent for the fluorine-based polymer and a good solvent for the plasticizer, and the boiling point is preferably lower than the melting point of the fluorine-based polymer film. Examples of such extraction solvents include hydrocarbons such as hexane and cyclohexane, halogenated hydrocarbons such as methylene chloride and 1,1,1-trichloroethane, alcohols such as ethanol and isopropanol, diethyl ether and tetrahydrofuran, and the like. Ethers, ketones such as acetone and 2-butanone, and water.

In the present invention, the first method for removing the plasticizer is to immerse the fluoropolymer film cut into a predetermined size in a container containing the extraction solvent and thoroughly wash it, and then air-dry the attached solvent. Or by drying with hot air. At this time, it is preferable to repeat the dipping operation and the washing operation many times since the plasticizer remaining in the fluoropolymer film is reduced. Moreover, in order to suppress shrinkage | contraction of a fluorine-type polymer film | membrane during a series of operation | movement of immersion, washing | cleaning, and drying, it is preferable to restrain the edge part of a fluorine-type polymer film | membrane.
The second method for removing the plasticizer is to continuously feed the fluoropolymer film into a tank filled with the extraction solvent and immerse it in the tank for a sufficient time to remove the plasticizer. Thereafter, the solvent adhered thereto is dried. At this time, the inside of the tank is divided into multiple stages, and the extraction solvent is supplied from the opposite direction to the running direction of the fluorine-based polymer film, or the multi-stage method in which the fluorine-based polymer film is sequentially fed to each tank having a concentration difference. It is preferable to apply a known means such as a counter-current method for providing a concentration gradient because the extraction efficiency is increased. In both the first and second methods, it is important to substantially remove the plasticizer from the fluoropolymer film. Substantially removing means removing the plasticizer in the fluoropolymer membrane to such an extent that the performance as a separation membrane is not impaired. The amount of the plasticizer remaining in the fluoropolymer membrane is 1% by weight. It is preferable that it is below, More preferably, it is 100 weight ppm or less. The amount of the plasticizer remaining in the fluorine polymer film can be quantified by gas chromatography, liquid chromatography, or the like. Further, it is preferable to heat the extraction solvent within the range of less than the boiling point of the solvent, preferably within the range of boiling point -5 ° C. or less, because the diffusion between the plasticizer and the solvent can be promoted.

  In the present invention, when the fluoropolymer film is subjected to heat treatment before, after, or before and after the step of removing the plasticizer, the shrinkage of the fluoropolymer film when the plasticizer is removed is reduced. The effects of improving the strength and heat resistance of the polymer film can be obtained. As a heat treatment method, a fluorine polymer film is placed in hot air, a fluorine polymer film is immersed in a heat medium, or a metal roll whose temperature is controlled by heating. There is a method in which a polymer film is brought into contact. In the heat treatment, it is preferable that the dimensions are fixed because it is possible to prevent blocking of fine holes. The temperature of the heat treatment is preferably performed below the melting point. In the case of polyvinylidene fluoride, the lower limit is 121 ° C. or higher, preferably 125 ° C. or lower, and the upper limit is 170 ° C. or lower, preferably 165 ° C. or lower. If the melting point is exceeded, problems such as breakage of the membrane and collapse of the pores may occur during the heat treatment.

A typical wet film forming method for obtaining the fluorine-based polymer film according to the present invention will be described.
The good solvent used in the wet film formation method for obtaining the fluorine-based polymer film according to the present invention is not limited as long as it is miscible with water, but 10 g or more can be dissolved in 100 g of pure water at 20 ° C. Further, it is preferable to dissolve 5% by weight or more of the fluorine-based polymer film as a film material. For example, N-methyl-2-pyrrolidone, N, N-dimethylacetamide, dimethyl sulfoxide, N, N-dimethylformamide, acetone, γ-butyrolactone and the like can be mentioned. From the viewpoint of danger, safety and toxicity, dimethyl sulfoxide, N, N-dimethylacetamide, and N-methyl-2-pyrrolidone are preferably used. These solvents can be used alone or in combination of two or more.

The membrane stock solution used in the wet film-forming method for obtaining the fluorine-based polymer film according to the present invention is not limited as long as a fluorine-based polymer film having the desired structure and performance can be produced. Regarding the concentration of the fluoropolymer in the membrane stock solution, the film formability improves as the concentration increases, but conversely, the porosity of the membrane decreases and the water permeability tends to decrease. Therefore, when the whole membrane stock solution is 100% by weight, the concentration range of the fluoropolymer varies depending on the molecular weight, but the lower limit is 2% by weight or more, preferably 5% by weight or more, particularly preferably 10% by weight or more. . Further, an upper limit of 50% by weight or less, preferably 40% by weight or less, particularly preferably 30% by weight or less, is preferably used.
In addition, the lower limit of the temperature of the membrane stock solution is preferably 0 ° C. or higher, preferably 10 ° C. or higher, particularly preferably 25 ° C. or higher, and the upper limit is a good solvent boiling point or lower in the membrane stock solution. If it is this temperature condition, the viscosity suitable for processing into a film | membrane preferable as a film | membrane stock solution can be obtained.
Addition of antioxidants, crystal nucleating agents, antistatic agents, flame retardants, lubricants, ultraviolet absorbers, etc., depending on the purpose, as long as the membrane undiluted solution used in the present invention does not affect the performance of the membrane to be produced. Mixing agents may be used.

In the case where a hydrophilic polymer is used in the wet film formation method for obtaining the fluorine-based polymer film according to the present invention, its role is mainly to promote and form the porous structure of the outer porous support layer part, The thickening effect of the stock solution is achieved. The amount of the hydrophilic polymer added to the membrane stock solution can be adjusted as appropriate in order to achieve stable film formation. When the viscosity of the film stock solution is low, film breakage or film breakage may occur at the time of film formation, and the film formability may become unstable. Conversely, if the viscosity of the membrane stock solution is too high, the porous support layer cannot be sufficiently grown, the porosity of the porous structure of the outer layer will be insufficient, and it will be difficult to obtain a membrane with the desired high permeability. . Furthermore, there is a concern that the stock solution discharged from the die may cause a melt fracture due to an increase in the viscosity of the membrane stock solution.
In the wet film formation method for obtaining the fluorine polymer film according to the present invention, the upper limit of the concentration of the hydrophilic polymer in the membrane stock solution is determined in accordance with the type and molecular weight of the hydrophilic polymer to be used. However, it is usually 40% by weight or less, preferably 30% by weight or less.

  The coagulating liquid used in the wet film-forming method for obtaining the fluorine-based polymer film according to the present invention is a substance that causes concentration-induced phase separation when in contact with the film stock solution and can form a film from the contact surface. There is no limitation. For example, pure water, a monoalcohol solvent, a polyol solvent, or a mixture of two or more of these is preferably used. Examples of the monoalcohol solvent include methanol, ethanol, propanol and the like. Examples of the polyol solvent include ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, glycerin, propylene glycol, 1,2-butanediol, 1,4-butanediol, and the like. In the coagulation liquid, polyvinyl alcohol, polyethylene glycol, polypropylene glycol, polyethylene oxide, polyethylene glycol-polypropylene glycol block copolymer, poly (meth) acrylamide, polyvinylpyrrolidone, polyhydroxyacrylate, polyhydroxymethacrylate, poly (meth) acrylic acid, Polymethacrylic acid, polyitaconic acid, polyfumaric acid, polycitraconic acid, poly-p-styrene sulfonic acid, sodium poly-p-styrene sulfonate, N, N-dimethyl (meth) acrylamide, carboxymethylcellulose, starch, corn starch, polychitosan It is also possible to add a water-soluble polymer such as polychitin. Although it depends on the molecular weight and the amount of the water-soluble polymer to be added, the filtration performance can be improved by adding these. The coagulation liquid may contain a good solvent such as N-methyl-2-pyrrolidone, N, N-dimethylacetamide, dimethyl sulfoxide, N, N-dimethylformamide, and γ-butyrolactone. In particular, when using a coagulation liquid containing a good solvent in a non-solvent, the composition differs depending on the composition of the membrane stock solution, the contact temperature between the membrane stock solution and the coagulation liquid, etc. In this case, 90% by weight or less is preferable as the weight% of the good solvent. Within this range, concentration-induced phase separation necessary and sufficient for forming a film can be sufficiently achieved.

  The film formation temperature in the wet film formation method for obtaining the fluoropolymer film according to the present invention is not limited as long as it is a temperature at which the film stock solution and the coagulation liquid are brought into contact with each other to cause concentration-induced phase separation. The lower limit of the film temperature is 0 ° C. or higher, preferably 10 ° C. or higher, particularly preferably 25 ° C. or higher. The upper limit is a temperature below each boiling point of the membrane stock solution or coagulation liquid, preferably a temperature lower by 5 ° C. or more from each boiling point, particularly preferably a temperature lower by 10 ° C. or more from the boiling point. For hollow fiber membranes, it depends on the temperature of the double nozzle. In a flat membrane, it is determined by the coagulation liquid temperature.

  Membrane stock solution and coagulation solution used in the wet film formation method for obtaining the fluorine-based polymer membrane according to the present invention, particularly coagulation solution (hereinafter referred to as internal coagulation solution) passing through the inside of the yarn during the production of the hollow fiber membrane, is dissolved after uniform dissolution. It is desirable to remove the gas. By removing the dissolved gas, film defects due to foaming of the dissolved gas can be remarkably improved. Further, by removing oxygen from the dissolved gas, the oxidation reaction to the material due to film processing at a high temperature is reduced. This step may be omitted when no gas is dissolved in the membrane stock solution, the coagulation solution, and the internal coagulation solution.

In the case of producing a hollow fiber membrane by a wet film forming method for obtaining a fluoropolymer membrane according to the present invention, a tank is provided directly below the spinning nozzle in order to further promote the coagulation by the membrane stock solution and the internal coagulating solution that has come out of the double spinning nozzle. (Hereinafter referred to as a coagulation tank) can be provided and brought into contact with a coagulation liquid (hereinafter referred to as an external coagulation liquid) filled in the coagulation tank.
When a hollow fiber membrane is produced by a wet film-forming method for obtaining a fluorine-based polymer membrane according to the present invention, in order to freely control the cross-sectional structure of the hollow fiber membrane not only to a uniform structure but also to various non-uniform structures. In addition, the distance from the spinneret to the external coagulation liquid surface (hereinafter referred to as idle running distance) and the temperature and humidity of the space from the spinneret to the external coagulation liquid surface can be adjusted. Although there is no limitation as long as the temperature and humidity of the space can be adjusted, for example, the lower limit of the free running distance is 0.001 m or more, preferably 0.005 m or more, particularly preferably 0.01 m or more, and the upper limit is 2.0 m or less, preferably Is 1.5 m or less, particularly preferably 1.2 m or less. The temperature in the space from the spinning nozzle to the external solidification surface is 10 ° C. or higher, preferably 20 ° C. or higher, particularly preferably 25 ° C. or higher as the lower limit. The humidity varies depending on the temperature, but the lower limit is 0% or more, preferably 10% or more, particularly preferably 30% or more, and the upper limit is 100% or less.

  The winding speed in the case of producing a hollow fiber membrane by a wet film-forming method for obtaining a fluorine-based polymer membrane according to the present invention includes various factors as production conditions, the shape of the spinning nozzle, the composition of the spinning dope, the internal coagulating liquid, Although the speed may vary depending on the composition of the external coagulation liquid, the stock solution, and the temperature of each coagulation liquid, a speed of approximately 300 m / hour to 9,000 m / hour is selected.

In the wet film-forming method for obtaining the fluorine-based polymer film according to the present invention, after coagulation with the coagulation liquid, the solvent removal can be promoted by immersing in a solvent removal tank in order to increase the strength of the film. The solvent removal liquid is a solvent that can remove the remaining solvent after concentration-induced phase separation by the coagulation liquid, and any solvent that does not dissolve the membrane can be used. In general, water, ethanol or the like is often used.
The drying temperature of the undried fluorinated polymer film of the present invention obtained by the wet film-forming method according to the present invention is not limited as long as it does not cause film breakage during drying. From the above, drying is performed within a temperature range below the melting temperature of the fluoropolymer. The drying temperature has a lower limit of 30 ° C. or higher, preferably 40 ° C. or higher, and the upper limit of 120 ° C. or lower, preferably 100 ° C. or lower. The time required for drying is determined by the relationship with the drying temperature, but is generally selected from 0.01 hours to 48 hours.

The olefin polymer film according to the present invention is mainly composed of an olefin polymer, but may contain other high molecular weight substances and additives as long as the characteristics of the olefin polymer are not impaired. It is also possible to carry out by combining two or more of these polymers.
The olefin polymer according to the present invention is a polymer synthesized using olefins or alkenes as monomers, and examples thereof include polyethylene, polypropylene, and poly-4-methyl 1-pentene. Furthermore, the said homopolymer and the said copolymer can also be mixed and used. Among these, polyethylene is preferable.

  The weight average molecular weight of the olefin polymer according to the present invention is preferably 50,000 or more, preferably 100,000 or more, particularly preferably 150,000 or more as a lower limit, and 5 million or less, preferably 2 million or less, particularly preferably as an upper limit. Is preferably 1 million or less. In general, resins having an average molecular weight exceeding 1,000,000 are difficult to measure by GPC, and as an alternative, the viscosity average molecular weight determined by the viscosity method can be assigned. If the average molecular weight is less than 50,000, the melt tension at the time of melt molding becomes small and the moldability is deteriorated, and the mechanical strength of the film is lowered. An average molecular weight exceeding 5 million is not preferable because uniform melt-kneading becomes difficult.

  In order to prevent clogging due to the adsorption of immunoglobulin, it is necessary to impart hydrophilicity to the membrane. Examples of the hydrophilization treatment include a method in which an olefin polymer film is immersed in a solution containing a surfactant and then dried to leave the surfactant in the olefin polymer film. A method of grafting a hydrophilic (meth) acrylic monomer or the like onto the pore surface of an olefin polymer film by irradiating the above-mentioned radiation or using a peroxide. And a method of immersing the olefin polymer film in a solution containing the hydrophilic polymer and then drying to form a hydrophilic polymer film on the pore surface of the olefin polymer film. Graft polymerization is most preferable in view of the persistence of the formation and the possibility of leakage of hydrophilic additives. In particular, the hydrophilic treatment by the radiation graft polymerization method disclosed in JP-A-62-2179540, JP-A-62-258711, and U.S. Pat. It is preferable in that a uniform hydrophilic layer can be formed on the surface of the pores.

  The hydrophilic monomer used in the graft polymerization of the present invention is not particularly limited as long as it is a hydrophilic monomer having a vinyl group. A monomer having one vinyl group is preferable. Furthermore, (meth) acrylic monomers containing a sulfone group, a carboxyl group, an amide group, a neutral hydroxyl group, a sulfonyl group, a sulfonic acid group, etc. can be suitably used, but when filtering a solution containing an immunoglobulin, A monomer containing a hydroxyl group is particularly preferred. The hydrophilic monomer according to the present invention is a monomer that is uniformly dissolved when mixed with 1% by volume of pure water at 25 ° C. under atmospheric pressure. For example, a vinyl monomer having a hydroxyl group such as hydroxypropyl acrylate, or a functional group serving as a precursor thereof, a vinyl monomer having an anion exchange group such as triethylammonium ethyl methacrylate, or a cation exchange group such as sulfopropyl methacrylate. And vinyl monomers having an amide bond such as vinyl pyrrolidone. Among them, a vinyl monomer having one or more hydroxyl groups or a functional group serving as a precursor thereof is preferable because the permeability of the immunoglobulin solution is the highest.

  Specifically, acrylic acid such as hydroxypropyl acrylate and 2-hydroxyethyl methacrylate or esters of methacrylic acid and polyhydric alcohols, alcohols having unsaturated bonds such as allyl alcohol, vinyl acetate, vinyl propionate, etc. Examples include enol esters. In addition, a hydrophilic monomer having one vinyl group and a crosslinking agent having two or more vinyl groups are used in a proportion of 20 mol% or more and 1,000 mol% or less with respect to the hydrophilic monomer. Hydrophilicity is sufficiently achieved by copolymerization by a polymerization method.

  The cross-linking agent used according to the present invention is a cross-linking agent having two or more vinyl groups that can be copolymerized with the hydrophilic monomer, and is introduced by contacting the membrane simultaneously with the hydrophilic monomer. The crosslinking agent preferably has a number average molecular weight of 200 or more and 2,000 or less, more preferably a number average molecular weight of 250 or more and 1,000 or less, and most preferably a number average molecular weight of 300 or more and 600 or less. When the number average molecular weight of the crosslinking agent is 200 or more and 2,000 or less, a high filtration rate of the immunoglobulin solution is obtained, which is preferable. In the present invention, any crosslinking agent having two or more vinyl groups can be used, but a hydrophilic crosslinking agent is preferred. Here, the hydrophilic cross-linking agent is a cross-linking agent that is uniformly dissolved when mixed with 1% by volume of pure water at 25 ° C. under atmospheric pressure.

  Specific examples of the crosslinking agent used in the present invention include divinylbenzene derivatives for aromatic systems, methacrylic acid crosslinking agents such as ethylene glycol dimethacrylate and polyethylene glycol dimethacrylate for aliphatic systems, ethylene glycol diacrylate, Examples include (meth) acrylic acid-based crosslinking agents such as polyethylene glycol diacrylate. A crosslinking agent having three reactive groups such as trimethylolpropane trimethacrylate can also be used. In addition, a mixture of two or more kinds of crosslinking agents can be used. In the present invention, it is most preferable to use polyethylene glycol dimethacrylate, polyethylene glycol diacrylate, or a mixture thereof from the viewpoints of immunoglobulin monomer permeability and immunoglobulin dimer removal performance.

  The graft polymerization method according to the present invention is not limited as long as it is a method for generating radicals. For example, radicals are generated in an olefin polymer film by means of addition of a radiation initiator, ionizing radiation, chemical reaction, or the like. Then, starting from the radical, the monomer is graft-polymerized to the film. In the present invention, any means can be employed for generating radicals in the olefin polymer film, but irradiation with ionizing radiation is preferable in order to generate uniform radicals throughout the film. As types of ionizing radiation, γ rays, electron beams, β rays, neutron rays, and the like can be used, but electron beams or γ rays are most preferable for implementation on an industrial scale. The ionizing radiation is obtained from radioactive isotopes such as cobalt 60, strontium 90, and cesium 137, or by an X-ray imaging apparatus, an electron beam accelerator, an ultraviolet irradiation apparatus, or the like.

  The irradiation dose of ionizing radiation according to the present invention is preferably from 1 kGy to 1,000 kGy. If it is less than 1 kGy, radicals are not generated uniformly, and if it exceeds 1,000 kGy, film strength may be reduced. Graft polymerization is generally divided into two types: a pre-irradiation method in which radicals are generated on a film and then contacted with a reactive compound, and a simultaneous irradiation method in which radicals are generated on the film in a state where the film is in contact with the reactive compound. Is done. In the present invention, any method can be applied, but the pre-irradiation method in which oligomer formation is small is most preferable.

  In the present invention, the contact between the olefin polymer film that has generated radicals, the hydrophilic monomer and the crosslinking agent is achieved in the liquid phase even in the gas phase, but in the present invention, the liquid phase in which the grafting reaction proceeds uniformly. The method of contacting with is a preferred method. In order to further promote the graft reaction more uniformly, it is desirable that the hydrophilic monomer and the crosslinking agent are dissolved in a solvent in advance and then contacted with the polymer olefin polymer membrane. The solvent for dissolving the hydrophilic monomer and the crosslinking agent is not particularly limited as long as it can be uniformly dissolved. Examples of such solvents include alcohols such as ethanol, isopropanol, and t-butyl alcohol, ethers such as diethyl ether and tetrahydrofuran, ketones such as acetone and 2-butanone, water, and mixtures thereof. .

In the graft polymerization according to the present invention, a reaction solution of 0.3% by volume to 30% by volume with a concentration of a hydrophilic monomer and a crosslinking agent is used, and 10 × 10 ^{−5 to} 100 with respect to 1 g of the olefin polymer membrane. It is desirable to carry out the reaction at a rate of × 10 ⁻⁵ m ³ . If graft polymerization is performed within this range, pores are not filled with the hydrophilic layer, and a film having excellent uniformity can be obtained.
The reaction temperature at the time of graft polymerization according to the present invention is not particularly limited as long as the polymerization reaction occurs, but it is generally carried out at 20 ° C. to 80 ° C.
In the graft polymerization according to the present invention, when the olefin polymer film and the hydrophilic monomer are brought into contact with each other, the hydrophilic monomer may be in any state of gas, liquid or solution, but forms a uniform hydrophilic layer. For this purpose, a liquid or a solution is preferable, and a solution is particularly preferable.

The hydrophilic olefin polymer membrane according to the present invention introduces an immunoglobulin monomer permeability and an immunoglobulin dimer by introducing a hydrophilic layer having a strong cross-linked structure into a hydrophobic olefin polymer membrane. The blocking property can be realized at a high level. Therefore, the crosslinking agent is used in a proportion of 20 mol% or more, preferably 30 mol% or more, and 1,000 mol% or less, preferably 500 mol% or less, more preferably 200 mol% or less as the upper limit as the lower limit with respect to the hydrophilic monomer. That is good.
In the present invention, a hydrophilic layer is introduced into a hydrophobic olefin polymer membrane to achieve high immunoglobulin monomer permeability. Therefore, the graft ratio grafted onto the hydrophobic olefin polymer membrane is preferably 3% or more, preferably 4% or more, more preferably 5% or more as the lower limit, and 50% or less, preferably 30% as the upper limit. Below, more preferably 20% or less. When the graft ratio is less than 3%, the hydrophilicity of the membrane is insufficient, causing a rapid decrease in the filtration rate accompanying protein adsorption. If it exceeds 50%, relatively small pores are filled with the hydrophilic layer, and a sufficient filtration rate cannot be obtained. The graft rate referred to here is a value defined by the following formula (24).
Graft rate (%) =
(Membrane weight after grafting−membrane weight before grafting) / membrane weight before grafting × 100 (24)

  The degree of hydrophilicity of the olefin polymer film according to the present invention can be evaluated by the contact angle. The average value of the advancing contact angle and the receding contact angle at 25 ° C. is preferably 60 degrees or less, more preferably 45 degrees or less, and still more preferably 30 degrees or less. In addition, as a simple evaluation method, when the olefin polymer membrane is brought into contact with water, it may be determined that the membrane has sufficient hydrophilicity if water spontaneously permeates into the pores of the membrane.

The molecular weight cut off of the olefin polymer membrane according to the present invention is only required to be able to sufficiently separate the immunoglobulin monomer and dimer, and the lower limit is 100,000 or more, preferably 150,000 or more, more preferably 250,000 or more. The upper limit should be set to less than 500,000, preferably 450,000 or less, and more preferably 400,000 or less. If the molecular weight cut-off is less than 100,000, there is a problem that the amount of immunoglobulin permeated decreases, and if it is more than 500,000, both the immunoglobulin monomer and dimer permeate through the membrane. Decreases.
The molecular weight cut-off according to the present invention includes proteins such as albumin (66,000), γ-globulin (160,000), catalase (232,000), ferritin (440,000), thyroglobulin (669,000), and PEG. Then, dead end filtration is performed using dextran or the like, and the molecular weight is calculated as the molecular weight at which the rejection is 90% from the relationship between the molecular weight and the rejection.

The method for producing the olefin polymer film according to the present invention is not limited in any way, and examples thereof include a melt film forming method. The melt film-forming method is a method in which a film material and a plasticizer are uniformly mixed by heating and then phase separation is generated by cooling, and a film is obtained by extracting the plasticizer from the obtained film film. It is.
A typical melt film forming method for obtaining an olefin polymer film according to the present invention includes the following steps (a) to (c).
(A) A step of heating a composition containing an olefin polymer and a plasticizer to a temperature equal to or higher than the crystal melting point of the olefin polymer and uniformly dissolving the composition, and then discharging the composition from a discharge port to form a film;
(B) A non-volatile liquid having partial solubility in the olefin polymer while the film is taken at a take-off speed such that the draft ratio defined in the following formula (25) is 1 or more and 15 or less. In a state where the temperature is heated to 100 ° C. or higher, and the other film surface is cooled. Draft ratio = (film take-off speed) / (discharge at composition discharge port) (Speed) (25)
(C) removing a substantial part of the plasticizer and the non-volatile liquid;

  The polymer concentration used in the melt film forming method for obtaining the olefin polymer film according to the present invention is preferably 20 to 90% by weight, more preferably 30 to 80% by weight in the composition containing the olefin polymer and the plasticizer. Most preferably, it is 35 to 70% by weight. When the polymer concentration is less than 20% by weight, the film formability deteriorates, and there are problems such as insufficient mechanical strength. If the polymer concentration exceeds 90% by weight, the pore size of the resulting olefin polymer membrane will be too small and the porosity will be small, so the filtration rate will be low and it will not be practical.

The plasticizer used in the melt film forming method for obtaining the olefin polymer film according to the present invention includes a homogeneous solution at a temperature equal to or higher than the crystal melting point of the resin when mixed with the olefin polymer in the composition for producing the olefin polymer film. A non-volatile solvent capable of forming is used. The non-volatile solvent mentioned here has a boiling point of 250 ° C. or higher under atmospheric pressure. The plasticizer may be liquid or solid at a room temperature of 20 ° C. Further, a so-called solid-liquid phase separation type plasticizer that has a heat-induced solid-liquid phase separation point at a temperature of room temperature or higher when a homogeneous solution with an olefin polymer is cooled may be used. Some plasticizers have a heat-induced liquid-liquid phase separation point at a temperature equal to or higher than normal temperature when a homogeneous solution with an olefin polymer is cooled. When an agent is used, the obtained olefin polymer membrane tends to have a large pore size. The plasticizer used here may be a single item or a mixture of a plurality of substances.
The method for measuring the heat-induced solid-liquid phase separation point is a thermal analysis such as differential scanning calorimetry (DSC) using a sample obtained by melt-kneading a composition of a predetermined concentration containing an olefin polymer and a plasticizer in advance. By measuring the exothermic peak temperature of the resin. Further, the method for measuring the crystallization point of the resin can be obtained by thermal analysis in the same manner using a sample obtained by previously kneading the resin as a sample.

As a plasticizer concerning this invention, the plasticizer currently disclosed by the international publication 01/28667 pamphlet is mentioned. That is, it is a plasticizer having a phase separation point depression constant α defined by the following formula (26) of 0 to 40 ° C., preferably 1 to 35 ° C., more preferably 5 to 30 ° C. It is an agent. When the phase separation point depression constant exceeds 40 ° C., the uniformity and strength of the pore diameter are lowered, which is not preferable.
α = 100 × (T _c ⁰ −T _c ) ÷ (100−C) (26)
α is the phase separation point drop constant (℃)
T _c ⁰ is the crystallization temperature of the olefin polymer (℃)
T _c is the thermally induced solid-liquid phase separation point (° C.) of the composition
C is the concentration of olefinic polymer in the composition (wt%)

Specifically, phthalic acid esters having an ester chain carbon chain length of 7 or less, adipic acid esters, sebacic acid esters, phosphate esters having an ester chain carbon chain length of 8 or less, citrate esters, etc. Can be preferably used, in particular diheptyl phthalate, dicyclohexyl phthalate, dibutyl phthalate, diethyl phthalate, dimethyl phthalate, dibutyl adipate, dibutyl sebacate, triphenyl phosphate, tricresyl phosphate, diphenyl cresyl phosphate, Tri (2-ethylhexyl) phosphate, tributyl phosphate, tributyl acetylcitrate and the like are particularly preferable.
In the composition used in the present invention, an antioxidant, a crystal nucleating agent, an antistatic agent, a flame retardant, a lubricant, an ultraviolet absorber and the like are added depending on the purpose as long as the performance of the film to be produced is not affected. Mixing agents may be used.

In the melt film-forming method for obtaining the olefin polymer film according to the present invention, the first method for uniformly dissolving the composition containing the olefin polymer and the plasticizer is a continuous resin kneading apparatus such as an extruder. In this method, a plasticizer is introduced at an arbitrary ratio while the resin is heated and melted, and screw kneading is performed to obtain a uniform solution. The form of the resin to be charged may be any of powder, granule, and pellet. Moreover, when making it melt | dissolve uniformly by such a method, it is preferable that the form of a plasticizer is a normal temperature liquid. As an extruder, a single screw type extruder, a biaxial different direction screw type extruder, a biaxial same direction screw type extruder, etc. can be used.
The second method for uniformly dissolving the composition containing the olefin polymer and the plasticizer is to mix and disperse the olefin polymer and the plasticizer in advance using a stirring device such as a Henschel mixer. This is a method of obtaining a uniform solution by putting a product into a continuous resin kneader such as an extruder and melt-kneading. The form of the composition to be added may be a slurry when the plasticizer is a liquid at room temperature, and may be a powder or a granule when the plasticizer is a solid at room temperature.
The third method of uniformly dissolving a composition containing an olefin polymer and a plasticizer is a method using a simple resin kneader such as a Brabender or a mill, or a method of melt kneading in another batch kneading container. is there. According to this method, since it is a batch type process, the productivity is not good, but there is an advantage that it is simple and highly flexible.

In the melt film-forming method for obtaining an olefin polymer film according to the present invention, a composition containing an olefin polymer and a plasticizer is heated and dissolved at a temperature equal to or higher than the crystal melting point of the olefin polymer, After the step (a) of extruding from the discharge port of the circular die and the annular nozzle into a flat membrane shape and a hollow fiber shape, the process proceeds to the step (b) in which it is cooled and solidified and molded. Form a structure.
In the melt film forming method for obtaining an olefin polymer film according to the present invention, a composition containing a uniformly heated and dissolved olefin polymer and a plasticizer is discharged from a discharge port, and is defined by the following formula (27). A non-volatile liquid having partial solubility with respect to the olefin polymer is brought into contact with the olefin polymer while the film is taken at a take-up speed such that the ratio is 1 or more and 15 or less, thereby forming a film.
Draft ratio = (film take-off speed) / (discharge speed at discharge port of composition) (27)

The draft ratio is preferably 1.5 or more as a lower limit, more preferably 2 or more, and preferably 10 or less, more preferably 7 or less as an upper limit. When the draft ratio is less than 1, no tension is applied to the film and the moldability is lowered. When the draft ratio exceeds 15, the film is stretched and it is difficult to form a sufficiently thick coarse structure layer.
The discharge rate of the composition at the discharge port is given by the following formula (28).
Discharge speed at the discharge port of the composition =
(Volume of composition discharged per unit time) / (area of discharge port) (28)

A preferable range of the discharge speed is 1 m / min or more, preferably 3 m / min or more as a lower limit, and 60 m / min or less, preferably 40 m / min or less as an upper limit. When the discharge speed is less than 1 m / min, in addition to a decrease in productivity, problems such as a large fluctuation in discharge amount occur. On the contrary, when the discharge speed exceeds 60 m / min, since the discharge amount is large, turbulent flow may occur at the discharge port, and the discharge state may become unstable. The take-up speed can be set according to the discharge speed, but the lower limit is 1 m / min or more, preferably 3 m / min or more, and the upper limit is 200 m / min or less, preferably 150 m / min or less. When the take-up speed is less than 1 m / min, productivity and moldability are deteriorated, and when the take-up speed exceeds 200 m / min, the cooling time is shortened, and the tension on the film is increased, so that the film is broken. It becomes easy to get up.

  In the present invention, an extraction solvent is used to remove the plasticizer. The extraction solvent is preferably a poor solvent for the olefin polymer and a good solvent for the plasticizer, and the boiling point is preferably lower than the melting point of the olefin polymer film. Examples of such extraction solvents include hydrocarbons such as hexane and cyclohexane, halogenated hydrocarbons such as methylene chloride and 1,1,1-trichloroethane, alcohols such as ethanol and isopropanol, diethyl ether and tetrahydrofuran, and the like. Ethers, ketones such as acetone and 2-butanone, and water.

In the present invention, the first method for removing the plasticizer is to immerse the olefin polymer film cut into a predetermined size in a container containing the extraction solvent and thoroughly wash it, and then air-dry the attached solvent. Or by drying with hot air. At this time, it is preferable to repeat the dipping operation and the washing operation many times because the plasticizer remaining in the olefin polymer film is reduced. Moreover, in order to suppress shrinkage | contraction of an olefin type polymer film during a series of operation | movement of immersion, washing | cleaning, and drying, it is preferable to restrain the edge part of an olefin type polymer film.
The second method for removing the plasticizer is to continuously feed the olefin polymer membrane into a tank filled with the extraction solvent and immerse it in the tank for a sufficient time to remove the plasticizer. Thereafter, the solvent adhered thereto is dried. At this time, by dividing the inside of the tank into multiple stages, the extraction solvent is supplied from the opposite direction to the running direction of the olefin polymer film, or the multi-stage method in which the olefin polymer film is sequentially fed to each tank having a concentration difference. It is preferable to apply a known means such as a counter-current method for providing a concentration gradient because the extraction efficiency is increased.

  In both the first and second methods, it is important to substantially remove the plasticizer from the olefin polymer membrane. Substantially removing means removing the plasticizer in the olefin polymer membrane to such an extent that the performance as a separation membrane is not impaired. The amount of the plasticizer remaining in the olefin polymer membrane is 1% by weight. It is preferable that it is below, More preferably, it is 100 weight ppm or less. The amount of the plasticizer remaining in the olefin polymer film can be quantified by gas chromatography, liquid chromatography, or the like. Further, it is preferable to heat the extraction solvent within the range of less than the boiling point of the solvent, preferably within the range of boiling point -5 ° C. or less, because the diffusion between the plasticizer and the solvent can be promoted.

  In the present invention, when the olefin polymer film is subjected to heat treatment before, after, or before and after the step of removing the plasticizer, the shrinkage of the olefin polymer film when the plasticizer is removed is reduced. The effects of improving the strength and heat resistance of the polymer film can be obtained. The heat treatment may be performed by placing an olefin polymer film in hot air, by immersing the olefin polymer film in a heat medium, or by heating and adjusting a metal roll, etc. There is a method in which a polymer film is brought into contact. In the heat treatment, it is preferable that the dimensions are fixed because it is possible to prevent blocking of fine holes. The temperature of the heat treatment is preferably performed below the melting point of the object and the olefin polymer. In the case of polyethylene, the lower limit of the drying temperature is 60 ° C or higher, preferably 65 ° C or higher, and the upper limit is 100 ° C or lower, preferably 95 ° C or lower. If the melting point is exceeded, problems such as breakage of the membrane and collapse of the pores may occur during the heat treatment. The time required for drying is determined by the relationship with the drying temperature, but is generally selected from 0.01 hours to 48 hours.

The vinyl alcohol polymer according to the present invention was copolymerized with polyvinyl alcohol or modified polyvinyl alcohol such as partially acetalized ethylene, propylene, vinyl pyrrolidone, vinyl chloride, vinyl fluoride, methyl methacrylate, acrylonitrile, itaconic acid and the like. Copolymers (including block copolymers and graft copolymers) and derivatives thereof. Among these, an ethylene-vinyl alcohol copolymer is preferable.
The lower limit of the saponification degree of the vinyl alcohol polymer according to the present invention is 80 mol% or more, preferably 85 mol% or more, and the upper limit is 100 mol% or less, preferably 95 mol% or less.
The polyvinyl alcohol content in the vinyl alcohol polymer chain according to the present invention is preferably at least 30% by weight, more preferably 50% by weight. If it is less than 30% by weight, it is not preferable because problems such as low hydrophilicity of the film occur.
The vinyl alcohol polymer film according to the present invention is mainly composed of a vinyl alcohol polymer, but contains other high molecular weight substances and additives as long as the characteristics of the vinyl alcohol polymer are not impaired. Also good. It is also possible to carry out by combining two or more of these polymers.

The weight average molecular weight of the vinyl alcohol polymer according to the present invention has a lower limit of 5,000 or more, preferably 10,000 or more, particularly preferably 50,000 or more, and an upper limit of 2 million or less, preferably 900,000 or less. Particularly preferred is 800,000 or less. In general, GPC measurement is difficult for a resin having an average molecular weight exceeding 1,000,000. Therefore, a viscosity average molecular weight by a viscosity method can be assigned as a substitute. An average molecular weight of less than 5,000 is not preferable because the mechanical strength of the film is lowered. An average molecular weight exceeding 2 million is not preferable because uniform melt kneading becomes difficult.
In the present invention, additional treatment may be performed within a range that does not adversely affect the vinyl alcohol polymer film. Examples of the additional treatment include cross-linking treatment and functional group introduction by chemical surface modification.

The molecular weight cut off of the vinyl alcohol polymer membrane according to the present invention is only required to sufficiently separate the immunoglobulin monomer and dimer, and the lower limit is 100,000 or more, preferably 150,000 or more, more preferably 250,000 or more. The upper limit is less than 500,000, preferably 450,000 or less, more preferably 400,000 or less. If the molecular weight cut-off is less than 100,000, there is a problem that the amount of immunoglobulin permeated decreases, and if it is more than 500,000, both the immunoglobulin monomer and dimer permeate through the membrane. Decreases.
The molecular weight cut-off according to the present invention includes proteins such as albumin (66,000), γ-globulin (160,000), catalase (232,000), ferritin (440,000), thyroglobulin (669,000), and PEG. Then, dead end filtration is performed using dextran or the like, and the molecular weight is calculated as the molecular weight at which the rejection is 90% from the relationship between the molecular weight and the rejection.

The method for producing the vinyl alcohol polymer film according to the present invention is not limited in any way, and examples thereof include a wet film forming method. The wet film-forming method is a method in which a membrane stock solution in which a membrane material is dissolved in a good solvent is brought into contact with a coagulation liquid composed of another solvent that is miscible with the good solvent in the membrane stock solution but is incompatible with the membrane material. In this method, concentration-induced phase separation is generated from the contact surface to obtain a membrane.
The good solvent used in the wet film-forming method for obtaining the vinyl alcohol polymer film according to the present invention is preferably a solvent capable of dissolving 5% by weight or more of the vinyl alcohol polymer as the film material. Water, alcohol, dimethyl sulfoxide N, N-dimethylformamide, N, N-dimethylacetamide, N-methyl-2-pyrrolidone and the like. These solvents can be used alone or in combination of two or more. Water is most preferable from an industrial viewpoint. In addition to the above composition, boric acid that promotes coagulation, a surfactant that improves film formation stability, an antifoaming agent, and the like may be added as appropriate.

  The membrane stock solution used in the wet film-forming method for obtaining the vinyl alcohol polymer film according to the present invention is not limited as long as a vinyl alcohol polymer film having the desired structure and performance can be produced. Usually, a vinyl alcohol polymer and a pore size forming agent dissolved in a common solvent are used. Regarding the concentration of the vinyl alcohol polymer in the membrane stock solution, the film formability improves as the concentration increases, but conversely, the porosity of the membrane decreases and the water permeability tends to decrease. Therefore, when the whole membrane stock solution is 100% by weight, the concentration range of the vinyl alcohol polymer varies depending on the molecular weight, but the lower limit is 2% by weight or more, preferably 5% by weight or more, particularly preferably 10% by weight or more. is there. Further, an upper limit of 50% by weight or less, preferably 40% by weight or less, particularly preferably 30% by weight or less, is preferably used.

The temperature of the membrane stock solution in the wet film forming method for obtaining the vinyl alcohol polymer membrane according to the present invention is 0 ° C. or more, preferably 10 ° C. or more, particularly preferably 25 ° C. or more as the lower limit, and the good solvent in the membrane stock solution as the upper limit. The boiling point or lower is preferably used. If it is this temperature condition, the viscosity suitable for processing into a film | membrane preferable as a film | membrane stock solution can be obtained.
Addition of antioxidants, crystal nucleating agents, antistatic agents, flame retardants, lubricants, ultraviolet absorbers, etc., depending on the purpose, as long as the membrane undiluted solution used in the present invention does not affect the performance of the membrane to be produced. Mixing agents may be used.

Examples of the pore forming agent used in the wet film forming method for obtaining the vinyl alcohol polymer film according to the present invention include polyethylene glycol, polypropylene glycol, tetraethylene glycol, triethylene glycol, ethylene having an average molecular weight of 200 to 4,000,000. Examples include glycols such as glycol, alcohols such as methanol, ethanol and propanol, polyhydric alcohols such as glycerin and butanediol, esters such as ethyl lactate and butyl lactate, etc., alone or in combination of two or more. It is done.
The addition amount of the pore-forming agent according to the present invention varies depending on the type of vinyl alcohol polymer and the type of pore-forming agent, but the addition amount should be such that the membrane stock solution has an upper critical solution point described later. preferable. The above upper critical eutectic point is the temperature at which the membrane undiluted solution becomes transparent and uniform at a high temperature and when the temperature of the undiluted solution is gradually lowered, the temperature changes from a transparent solution to a cloudy solution. Synonymous with cloud point.

When a hydrophilic polymer is used in a wet film forming method for obtaining a vinyl alcohol polymer film according to the present invention, its role is mainly to promote and form the porous structure of the outer porous support layer portion, It has the effect of thickening the membrane stock solution. The amount of the hydrophilic polymer added to the membrane stock solution can be adjusted as appropriate in order to achieve stable film formation. When the viscosity of the film stock solution is low, film breakage or film breakage may occur at the time of film formation, and the film formability may become unstable. Conversely, if the viscosity of the membrane stock solution is too high, the porous support layer cannot be sufficiently grown, the porosity of the porous structure of the outer layer will be insufficient, and it will be difficult to obtain a membrane with the desired high permeability. . Furthermore, there is a concern that the stock solution discharged from the die may cause a melt fracture due to an increase in the viscosity of the membrane stock solution.
In the wet film-forming method for obtaining the vinyl alcohol polymer film according to the present invention, the upper limit of the concentration of the hydrophilic polymer in the membrane stock solution is determined according to the type and molecular weight of the hydrophilic polymer to be used. However, it is usually 40% by weight or less, preferably 30% by weight or less.

  The coagulating liquid used in the wet film formation method for obtaining the vinyl alcohol polymer film according to the present invention is a substance that can cause concentration-induced phase separation when in contact with the film stock solution and form a film from the contact surface. For example, the water-based coagulant can be exemplified by pure water, an aqueous solution of a dehydrating salt such as sodium sulfate, an aqueous solution of an alkaline substance such as sodium hydroxide or aqueous ammonia, and the like. It can also be used or may be used in combination. In addition to the water-based coagulant, for example, methanol, ethanol, propanol and the like can be mentioned. Examples of polyol solvents include vinyl alcohol polymers such as ethylene glycol, diethylene glycol, triethylene glycol, tetraethylene glycol, glycerin, propylene glycol, 1,2-butanediol, 1,4-butanediol, and the like. It is free to use an organic coagulant having a coagulation ability of water or in combination with water. In the coagulation liquid, polyvinyl alcohol, polyethylene glycol, polypropylene glycol, polyethylene oxide, polyethylene glycol-polypropylene glycol block copolymer, polyvinyl alcohol amide, polyvinyl pyrrolidone, polyhydroxy acrylate, polyhydroxy methacrylate, polyvinyl alcohol acid, polymethacrylic acid, polyitacon Water-soluble acids, polyfumaric acid, polycitraconic acid, poly-p-styrene sulfonic acid, sodium poly-p-styrene sulfonate, N, N-dimethylvinyl alcohol amide, carboxymethyl cellulose, starch, corn starch, polychitosan, polychitin It is also possible to add a polymer. Although it depends on the molecular weight and the amount of the water-soluble polymer to be added, the filtration performance can be improved by adding these. The coagulation liquid may contain a good solvent such as N-methyl-2-pyrrolidone, N, N-dimethylacetamide, dimethyl sulfoxide, N, N-dimethylformamide, and γ-butyrolactone. In particular, when using a coagulation liquid containing a good solvent in a non-solvent, the composition differs depending on the composition of the membrane stock solution, the contact temperature between the membrane stock solution and the coagulation liquid, etc. In this case, 90% by weight or less is preferable as the weight% of the good solvent. Within this range, concentration-induced phase separation necessary and sufficient for forming a film can be sufficiently achieved.

  In the wet film forming method for obtaining the vinyl alcohol polymer film according to the present invention, the coagulating liquid (hereinafter referred to as the internal coagulating liquid) that passes through the inside of the yarn during the production of the hollow fiber membrane is the same solution as the above external coagulating liquid. Alternatively, an organic solvent that has no coagulation ability with respect to a vinyl alcohol polymer such as hexane or liquid paraffin and is not miscible with the solvent of the membrane stock solution may be used. Alternatively, a dry / wet film forming method in which a gas such as air, nitrogen, or ammonia gas is introduced may be used.

  The film formation temperature in the wet film formation method for obtaining the vinyl alcohol polymer film according to the present invention is not limited as long as it is a temperature at which the film stock solution and the coagulation liquid are brought into contact with each other to cause concentration-induced phase separation. The lower limit of the film formation temperature is 0 ° C. or higher, preferably 10 ° C. or higher, particularly preferably 25 ° C. or higher. The upper limit is a temperature below each boiling point of the membrane stock solution or coagulation liquid, preferably a temperature lower by 5 ° C. or more from each boiling point, particularly preferably a temperature lower by 10 ° C. or more from the boiling point. For hollow fiber membranes, it depends on the temperature of the double nozzle. In a flat membrane, it is determined by the coagulation liquid temperature.

It is desirable to remove the dissolved gas after uniform dissolution of the membrane stock solution used in the wet film formation method for obtaining the vinyl alcohol polymer membrane according to the present invention, the coagulation solution, and the coagulation solution passing through the inside of the fiber during the production of the hollow fiber membrane. . By removing the dissolved gas, the defect of the film due to foaming of the dissolved gas can be remarkably improved. In addition, by removing oxygen from the dissolved gas, the oxidation reaction to the material due to film processing at a high temperature is reduced. This step may be omitted when no gas is dissolved in the membrane stock solution, the coagulation solution, and the internal coagulation solution. In addition, when a gas such as air, nitrogen, or ammonia gas is used as a coagulant as a dry / wet film forming method, this step is not performed.
When producing a hollow fiber membrane by a wet film formation method for obtaining a vinyl alcohol polymer membrane according to the present invention, in order to further promote the coagulation by the membrane stock solution and the internal coagulation solution from the double spinning nozzle, A tank (hereinafter referred to as a coagulation tank) can be provided and brought into contact with a coagulation liquid (hereinafter referred to as an external coagulation liquid) filled in the coagulation tank.
When producing a hollow fiber membrane by a wet film forming method for obtaining a vinyl alcohol polymer membrane according to the present invention, the structure of the hollow fiber membrane is freely controlled not only to a uniform structure but also to various non-uniform structures. Therefore, it is possible to adjust the distance from the spinning nozzle to the external coagulation liquid surface (hereinafter referred to as idle running distance) and the temperature and humidity of the space from the spinning nozzle to the external coagulation liquid surface. Although there is no limitation as long as the temperature and humidity of the space can be adjusted, for example, the lower limit of the free running distance is 0.001 m or more, preferably 0.005 m or more, particularly preferably 0.01 m or more, and the upper limit is 2.0 m or less, preferably Is 1.5 m or less, particularly preferably 1.2 m or less. The temperature in the space from the spinning nozzle to the external solidification surface is 10 ° C. or higher, preferably 20 ° C. or higher, particularly preferably 25 ° C. or higher as the lower limit. The humidity varies depending on the temperature, but the lower limit is 0% or more, preferably 10% or more, particularly preferably 30% or more, and the upper limit is 100% or less.

  The winding speed in the case of producing a hollow fiber membrane by the wet film-forming method according to the present invention is as follows: various factors as production conditions, the shape of the nozzle, the composition of the spinning stock solution, the composition of the internal coagulating liquid and the external coagulating liquid, and the stock solution Although the speed may vary depending on the temperature of each coagulating liquid, a speed of approximately 300 m / hour to 9,000 m / hour is selected.

In the wet film formation method for obtaining the vinyl alcohol polymer film according to the present invention, after coagulation with the coagulation liquid, the film may be subjected to treatment such as stretching, neutralization, washing with water, wet heat treatment, ammonium sulfate replacement, and drying as necessary. it can. In order to increase the strength of the membrane, it can be immersed in a solvent removal tank to promote solvent removal. The solvent removal liquid is a solvent that can remove the remaining solvent after concentration-induced phase separation by the coagulation liquid, and any solvent that does not dissolve the membrane can be used. In general, water, ethanol or the like is often used. Furthermore, it is modified by acetalization with monoaldehyde and / or polyhydric aldehyde such as formaldehyde, glutaraldehyde, benzaldehyde, glyoxal, and nonane dial, esterification, etherification, etc., and cross-linking with methylol compound or polyisocyanate It is possible to perform the conversion process alone or in combination. In addition, it can be subjected to heat stretching and / or heat treatment after spinning, and further to the above-described various modification treatments after heat stretching and / or heat treatment.
The drying temperature of the undried vinyl alcohol polymer film of the present invention obtained by the wet film-forming method according to the present invention is not limited as long as it does not cause film breakage during drying. Drying is performed within a temperature range of from 0 ° C to the melting temperature of the vinyl alcohol polymer. A preferable drying temperature is 30 ° C. or higher and 80 ° C. or lower. The time required for drying is determined by the relationship with the drying temperature, but is generally selected from 0.01 hours to 48 hours.

  The degree of hydrophilicity of the vinyl alcohol polymer film according to the present invention can be evaluated by the contact angle. The average value of the advancing contact angle and the receding contact angle at 25 ° C. is preferably 60 degrees or less, more preferably 45 degrees or less, and still more preferably 30 degrees or less. Further, as a simple evaluation method, when the vinyl alcohol polymer membrane is brought into contact with water, it may be determined that it has sufficient hydrophilicity if water spontaneously permeates into the pores of the membrane.

The cellulosic polymer membrane according to the present invention is mainly composed of cellulosic polymer, but may contain other high molecular weight substances and additives as long as the characteristics of the cellulosic polymer are not impaired. It is also possible to carry out by combining two or more of these polymers.
Cellulose polymers according to the present invention include cellulose ester compounds such as copper ammonia regenerated cellulose, cellulose diacetate, cellulose triacetate, cellulose propionate, cellulose butyrate, and cellulose phenylcarbanilate, and cellulose ethers such as methylcellulose and ethylcellulose. , And blend compounds combining these. Among them, copper ammonia regenerated cellulose is preferable.

  The weight average molecular weight of the cellulosic polymer according to the present invention is 5,000 or more, preferably 10,000 or more, particularly preferably 50,000 or more as a lower limit, and 1,000,000 or less, preferably 900,000 or less, especially as an upper limit. Preferably it is 800,000 or less. Within this range, sufficient strength and film formability can be obtained.

The molecular weight cutoff of the cellulose polymer membrane according to the present invention is only required to sufficiently separate the immunoglobulin monomer and dimer, and the lower limit is 100,000 or more, preferably 150,000 or more, more preferably 250,000 or more. The upper limit is less than 500,000, preferably 450,000 or less, more preferably 400,000 or less. If the molecular weight cut-off is less than 100,000, there is a problem that the amount of immunoglobulin permeated decreases, and if it is more than 500,000, both the immunoglobulin monomer and dimer permeate through the membrane. Decreases.
The molecular weight cut-off according to the present invention includes proteins such as albumin (66,000), γ-globulin (160,000), catalase (232,000), ferritin (440,000), thyroglobulin (669,000), and PEG. Then, dead end filtration is performed using dextran or the like, and the molecular weight is calculated as the molecular weight at which the rejection is 90% from the relationship between the molecular weight and the rejection.

  The method for producing the hollow fiber of the cellulosic polymer membrane according to the present invention is not limited in any way. For example, the spinning dope is supplied from the outer spinning outlet of the annular double spinning nozzle, and the above-mentioned spinning solution is fed from the central spinning outlet of the annular double spinning nozzle. The internal coagulation liquid, which is a microphase separation and coagulation liquid for the spinning dope, is simultaneously discharged and introduced into the spinning cylinder. A spinning cylinder is a cylinder directly connected to a spinning nozzle. The spinning cylinder is filled with an external liquid to be brought into contact with the external coagulation liquid immediately after the spinning raw liquid is discharged, and is constantly fed, and flows down in the descending pipe together with the spinning raw liquid. At this time, particles are formed by microphase separation, and the three-dimensionally connected membrane structure is fixed to complete the porous membrane structure.

The good solvent used in the wet film-forming method for obtaining the cellulose polymer film according to the present invention is not limited as long as it dissolves the cellulose polymer, and examples thereof include a copper ammonia solution.
The membrane stock solution used in the wet film-forming method for obtaining the cellulose polymer film according to the present invention is not particularly limited as long as a cellulose polymer film having a target structure and performance can be produced. Regarding the concentration of the cellulosic polymer in the membrane stock solution, the film formability improves as the concentration increases, but conversely, the porosity of the membrane decreases and the water permeability tends to decrease. Therefore, when the whole membrane stock solution is 100% by weight, the concentration range of the cellulosic polymer varies depending on the molecular weight, but the lower limit is 2% by weight or more, preferably 5% by weight or more, particularly preferably 10% by weight or more. . Moreover, as an upper limit, the solution which melt | dissolved uniformly in 25 weight% or less, Preferably it is 20 weight% or less, Most preferably, it is 15 weight% or less is used suitably. When the cellulose concentration is less than 2% by weight, the resulting hollow fiber membrane has insufficient mechanical properties, and when it exceeds 25% by weight, the spinning solution adjustment and spinning operation become difficult.
The lower limit of the temperature of the membrane stock solution according to the present invention is preferably 0 ° C. or higher, preferably 10 ° C. or higher, particularly preferably 25 ° C. or higher, and the upper limit is a good solvent boiling point or lower in the membrane stock solution. If it is this temperature condition, the viscosity suitable for processing into a film | membrane preferable as a film | membrane stock solution can be obtained.
The film stock solution according to the present invention includes additives such as an antioxidant, a crystal nucleating agent, an antistatic agent, a flame retardant, a lubricant, and an ultraviolet absorber as long as the performance of the film to be produced is not affected. Can be mixed.

The coagulating liquid used in the wet film-forming method for obtaining the cellulose polymer film according to the present invention is a substance that can cause concentration-induced phase separation when in contact with the membrane stock solution and can form a film from the contact surface. Although it does not limit at all, for example, pure water, water, perchlene, trichlene, trichlorotrifluoroethane, methanol, ethanol, propanol, acetone, methyl ethyl ketone, sodium hydroxide, sulfuric acid, ammonium sulfate, formic acid, acetic acid, propionic acid, glycerin, polyethylene glycol Examples thereof include a liquid that exhibits non-coagulability or fine coagulation with respect to the spinning solution. Such a coagulant is appropriately selected depending on the type of spinning solution. A solution containing at least one selected from these coagulating liquids or a mixed liquid thereof is preferably used. Preferably, a mixed solution composed of acetone, ammonia and water is good.
The internal coagulating liquid and the external coagulating liquid used in the wet film forming method for obtaining the cellulose polymer film according to the present invention may be the same coagulating liquid or different coagulating liquids.

  As a film forming method for obtaining a cellulose polymer film according to the present invention, dry, wet using gas such as air, nitrogen, carbon dioxide, argon, oxygen, so-called Freon gas such as tetrafluoromethane, hexafluoroethane, and other halogen gas. You may manufacture by the film-forming method.

  The film formation temperature in the wet film formation method for obtaining the cellulose polymer film according to the present invention is not limited as long as it is a temperature at which the membrane stock solution and the coagulation liquid are brought into contact with each other to cause concentration-induced phase separation. The lower limit of the film temperature is 0 ° C. or higher, preferably 10 ° C. or higher, particularly preferably 25 ° C. or higher. The upper limit is a temperature below each boiling point of the membrane stock solution or coagulation liquid, preferably a temperature lower by 5 ° C. or more from each boiling point, particularly preferably a temperature lower by 10 ° C. or more from the boiling point. For hollow fiber membranes, it depends on the temperature of the double nozzle. In a flat membrane, it is determined by the coagulation liquid temperature.

  The membrane stock solution used in the wet film forming method for obtaining the cellulose polymer membrane according to the present invention, the coagulating solution, in particular, the internal coagulating solution passing through the inside of the yarn during the production of the hollow fiber membrane can be dissolved uniformly and then the dissolved gas can be removed. desirable. By removing the dissolved gas, film defects due to foaming of the dissolved gas can be remarkably improved. Further, by removing oxygen from the dissolved gas, the oxidation reaction to the material due to film processing at a high temperature is reduced. This step may be omitted when no gas is dissolved in the membrane stock solution, the coagulation solution, and the internal coagulation solution. In addition, when a gas such as air, nitrogen, ammonia gas or the like is used as a coagulant as a dry / wet film forming method, this step is not performed.

In the wet film forming method for obtaining the cellulose polymer film according to the present invention, a distance from the spinning nozzle to the external coagulation liquid surface (hereinafter referred to as an idle running distance) is provided and introduced into the external coagulation liquid, or directly external coagulation. It may be introduced into the liquid. The free running distance is preferably such a length that the spinning dope enters the external coagulation liquid straight. For example, it is 0.5 m or less, preferably 0.2 m or less, particularly preferably 0.1 m or less. If the free running distance is long, the moldability is deteriorated and the hollow fiber shape cannot be maintained.
The temperature in the space of the free running distance according to the present invention is 10 ° C. or higher, preferably 20 ° C. or higher, particularly preferably 25 ° C. or higher as the lower limit. The humidity varies depending on the temperature, but the lower limit is 0% or more, preferably 10% or more, particularly preferably 30% or more, and the upper limit is 100% or less.
The spinning dope has a shape of a hollow fiber membrane when it flows down the descending tube, and this hollow fiber membrane is drawn out from the opening of the ascending tube and wound around the winding frame.

The winding speed in the case of producing a hollow fiber membrane by a wet film-forming method for obtaining a cellulose polymer membrane according to the present invention is as follows: various factors that are production conditions, the shape of the spinneret, the composition of the spinning dope, the internal coagulating liquid, Although it may change depending on the composition of the external coagulation liquid, the temperature of the raw liquid and each coagulation liquid, etc., the lower limit is 100 m / hour or more, more preferably 200 m / hour or more, and the upper limit is 1,000 m / hour or less, More preferably, it is 500 m / hour or less.
In the wet film forming method for obtaining the cellulose polymer film according to the present invention, after the coagulation with the coagulating liquid, the desolvation can be promoted by immersing in a solvent removal tank in order to increase the strength of the film. The solvent removal liquid is a solvent that can remove the remaining solvent after concentration-induced phase separation by the coagulation liquid, and any solvent that does not dissolve the membrane can be used. In general, water, ethanol or the like is often used.

  The drying temperature of the undried cellulosic polymer film obtained by the wet film-forming method according to the present invention is not limited as long as it does not cause film breakage during drying. Drying is performed within a temperature range below the melting temperature of the polymer. The lower limit of the drying temperature is 30 ° C or higher, more preferably 40 ° C or higher, and the upper limit is 80 ° C or lower, more preferably 70 ° C or lower. The time required for drying is determined depending on the relationship with the drying temperature, but is generally from 0.01 to 48 hours. Further, after scouring with water and an aqueous inorganic salt solution, a known membrane pore size retaining agent such as glycerin or polyethylene glycol may be applied and dried.

When the flow rate of the external coagulating liquid in the spinning cylinder according to the present invention (external coagulating liquid speed) is extremely slower than the winding speed, it becomes undesirably extending in the spinning cylinder. In the present invention, the relationship between the external coagulation liquid speed and the winding speed is preferably such that the speed difference is 20% or less.
The external coagulation liquid speed according to the present invention is preferably high in order to suppress the bath resistance in the coagulation bath. However, if the flow rate of the external liquid is too high, the yarn swinging of the hollow fiber membrane becomes intense and spinning becomes difficult, so it is necessary to set it to an appropriate value. Most preferably, the maximum flow rate is selected in such a range that the yarn does not sway in the hollow fiber membrane.

The larger the diameter of the spinning cylinder according to the present invention, the easier the spinning operation, but a smaller one is desirable because a large amount of coagulation liquid is required, and the lower limit is 3 mm or more, more preferably 5 mm. The upper limit is good, and the upper limit is 20 mm or less, more preferably 10 mm or less.
The length of the spinning tube according to the present invention must be capable of providing an appropriate coagulation time corresponding to the formation of the hollow fiber membrane structure. It is preferable to be set to this. Any material can be used as long as it is durable to the coagulating liquid, but a transparent material that can observe the spinning state is desirable, for example, glass, polyethylene. Polypropylene, polytetrafluoroethylene, etc. can be used. Among them, polytetrafluoroethylene having the feature that the spinning operation is easy because the spinning stock solution is difficult to adhere is the most preferable material.

The thickness of the ultrafiltration membrane of the present invention is 15 μm or more, preferably 20 μm or more as the lower limit, and 2,000 μm or less, preferably 1,000 μm or less, particularly preferably 500 μm or less as the upper limit. If the film thickness is less than 15 μm, the strength of the ultrafiltration membrane tends to be insufficient, such being undesirable. On the other hand, if it exceeds 2,000 μm, there is a tendency that the permeation performance of the immunoglobulin monomer is insufficient, which is not preferable.
When the ultrafiltration membrane according to the present invention has a dense layer on the inner surface of the hollow fiber or on one side of the flat membrane, the thickness of the dense layer is usually in order to improve the permeation of the immunoglobulin solution. It is 100 μm or less, preferably 10 μm or less, and more preferably 1 μm or less.

  The porosity of the ultrafiltration membrane according to the present invention has a lower limit of 30% or more, preferably 40% or more, particularly preferably 50%, and an upper limit of 95% or less, preferably 90% or less, particularly preferably. Is preferably 85% or less. If the porosity is less than 30%, the filtration rate becomes insufficient, and if it exceeds 95%, the strength of the ultrafiltration membrane becomes insufficient. The porosity is a numerical value obtained from the apparent volume obtained from the cross-sectional area and length of the membrane, the weight of the membrane, and the true density of the membrane material.

The shape of the ultrafiltration membrane according to the present invention is not particularly limited as long as the fractionation performance can be expressed.For example, various shapes such as a hollow fiber shape, a flat membrane shape, and a tube shape can be used. A hollow fiber shape having a large filtration effective membrane area compared to the volume is effective.
The membrane surface structure of the ultrafiltration membrane according to the present invention is not particularly limited, and examples thereof include single holes such as a circle and an ellipse, continuous holes continuously connected, net-like fine holes, slit-like fine holes, and the like.

  In the present invention, the ultrafiltration membrane may be an ultrafiltration membrane as long as the surface of the membrane in contact with the immunoglobulin is used, and a substrate (support) made of any material may be used in order to maintain the structure. For example, a woven fabric, a nonwoven fabric, a porous inorganic material or the like is used as another base material (support) to increase physical strength, and a membrane obtained by molding an ultrafiltration membrane on these base materials.

  The apparatus for performing cross flow filtration according to the present invention is not limited as long as it can control the immunoglobulin concentration, linear velocity, pressure, etc. For example, the concentration of the immunoglobulin solution is monitored with an absorptiometer, A cross-flow filtration device that integrates a device for supplying a diluent to keep the concentration of globulin solution constant and a device for controlling the tangential line speed and the pressure across the ultrafiltration membrane with respect to the ultrafiltration membrane. Is mentioned.

Specifically, a cross flow filtration device as shown in FIG. The concentration of the solution in the immunoglobulin source liquid tank (4) is monitored by a concentration controller (11) incorporating an absorptiometer, and the signal is sent to the liquid feeding pump 1 (2) to control rotation and dilution. While adding the diluent in the liquid tank (1), the solution concentration in the immunoglobulin source liquid tank (4) is controlled. Furthermore, pressure and flow rate are monitored by pressure gauge 1 (5), pressure gauge 2 (6), and flow meter (10). From pressure / flow rate controller (12), adjustment valve (7) and feed pump 2 (3) are monitored. To control the ultrafiltration membrane module (8) so that the linear velocity in the tangential direction and the pressure across the ultrafiltration membrane become set values. An apparatus capable of measuring the concentration of the immunoglobulin permeate in the obtained immunoglobulin permeate tank (9) and the ratio of the immunoglobulin monomer to the immunoglobulin dimer, such as an absorptiometer or GPC It may be connected to.
The “immunoglobulin original solution” according to the present invention is an immunoglobulin solution used for fractionation performance evaluation and separation. Further, the “immunoglobulin permeate” is a solution separated and permeated by an “ultrafiltration membrane”.

  Further, a cross flow filtration device as shown in FIG. 2 may be used. A device capable of monitoring the concentration between the immunoglobulin source solution tank (4) and the ultrafiltration membrane module (8), for example, a UV flow cell (13) is provided, and the solution concentration in the immunoglobulin source solution tank (4) Is monitored by the concentration controller (11), and the signal is sent to the liquid delivery pump 1 (2) to control the rotation, and while adding the diluent in the diluent tank (1), the original immunoglobulin tank (4) The concentration of the solution in is controlled. Furthermore, pressure and flow rate are monitored with pressure gauge 1 (5), pressure gauge 2 (6), and flow meter (10). From pressure / flow rate controller (12), adjustment valve (7) and feed pump 2 (3) are monitored. To control the ultrafiltration membrane module (8) so that the linear velocity in the tangential direction and the pressure across the ultrafiltration membrane become set values. A device capable of measuring the concentration of the immunoglobulin permeate in the immunoglobulin permeate tank (9) and the ratio of the immunoglobulin monomer and dimer, for example, an absorptiometer or GPC is connected to the present crossflow filtration device. You may do it.

Addition of anti-aggregation agents, stabilizers, preservatives, etc. to the immunoglobulin solution as long as it does not affect the ability to reduce the permeation of proteins with a lower isoelectric point than immunoglobulin without causing denaturation or aggregation of the immunoglobulin. You may do it. Depending on the type and concentration of the aggregation inhibitor, the electrical properties in the immunoglobulin may be reduced, affecting the permeability of proteins having a lower isoelectric point than the immunoglobulin.
During filtration, the immunoglobulin is stressed and may aggregate and become cloudy. In that case, there is a method of reducing by adding a surfactant or saccharide as an aggregation inhibitor.

  The surfactant according to the present invention is not limited in any way as long as it does not cause denaturation or aggregation of immunoglobulin and does not affect the permeation of proteins having a lower isoelectric point than immunoglobulin. Examples thereof include an activator, a nonionic surfactant, a cationic surfactant, and an anionic surfactant.

  Examples of the amphoteric surfactant according to the present invention include amino acids, amino acid derivatives, sodium alkylamino fatty acids, alkylbetaines, and alkylamine oxides. Among these, amino acids and / or amino acid derivatives are particularly preferable. Amino acids are substances that exhibit the effect of suppressing the aggregation of immunoglobulins, and are not limited as long as they do not affect immunoglobulin denaturation and can reduce the permeation of proteins having a lower isoelectric point than immunoglobulins. Lysine, arginine, alanine, cysteine, glycine, serine, proline and the like. Among them, lysine, arginine and alanine are preferable, and lysine exhibits an effect in particular. The amino acid derivative according to the present invention is a substance obtained by chemically modifying an amino acid, and includes an acetylated amino acid and an acylated amino acid. The amino acid and / or the amino acid derivative may be used in the form of an acid addition salt. Examples of the acid that can form an acid addition salt include hydrochloric acid and sulfuric acid. Two or more of these can be used in combination. Further, it can be used in combination with other aggregation inhibitors.

  Examples of the nonionic surfactant according to the present invention include polyoxyethylene alkyl ether, polyoxyethylene alkylphenyl ether, sucrose fatty acid ester, sorbitan fatty acid ester, polyoxyethylene sorbitan fatty acid ester, and fatty acid alkanolamide. . Of these, polyoxyethylene alkyl ether polymers and / or derivatives thereof are particularly preferable. Polyoxyethylene alkyl ether is a substance that exhibits the effect of suppressing the aggregation of immunoglobulins, and does not cause the denaturation of immunoglobulins. The type is not particularly limited, and examples thereof include polyethylene glycol and / or polyethylene glycol derivatives, and include a polyethylene glycol, polyethylene oxide, polyethylene glycol-polypropylene glycol block copolymer, and a surface activity containing polyethylene glycol as a hydrophilic segment. An agent, a block copolymer, and a graft copolymer can be sufficiently used as an aggregation inhibitor. Two or more of these can be used in combination. Further, it can be used in combination with other aggregation inhibitors. Two or more of these can be used in combination. Further, it can be used in combination with other aggregation inhibitors.

Examples of the cationic surfactant according to the present invention include alkyltrimethylammonium salts and dialkyldimethylammonium salts.
Examples of the anionic surfactant according to the present invention include fatty acid sodium, fatty acid potassium, sodium alphasulfo fatty acid ester, linear sodium alkylbenzene sulfonate, sodium alkyl sulfate, sodium alkyl ether sulfate, sodium alpha olefin sulfonate, Examples include sodium alkyl sulfonate.

  The lower limit value of the molecular weight of the surfactant according to the present invention is 30 Da or more, more preferably 50 Da or more, and the upper limit value is 50,000 Da or less, more preferably 30,000 Da or less. If it is less than 30 Da, a sufficient aggregation-inhibiting effect cannot be obtained. Conversely, if it exceeds 50,000 Da, a surfactant or a complex of immunoglobulin and surfactant may cause clogging or fouling in the membrane. is there.

  The saccharides according to the present invention are substances that exhibit the effect of suppressing the aggregation of immunoglobulins, and do not affect the denaturation of immunoglobulins and can reduce the permeation of proteins having a lower isoelectric point than immunoglobulins. Although the type is not particularly limited, examples of specific sugars include glucose, sorbitol, and sucrose. In addition to the aggregation suppressing effect, the surfactants, saccharides, and the like have effects such as prevention of immunoglobulin deterioration and adsorption.

  The concentration of the aggregation inhibitor in the present invention depends on the type of the aggregation inhibitor, but the lower limit is preferably 0.01 g / L or more, more preferably 0.05 g / L or more, and the upper limit. Is preferably 200 g / L or less, more preferably 150 g / L or less. When the lower limit value is less than 0.01 g / L, the effect of suppressing the aggregation of immunoglobulin is low, and when the upper limit value is more than 200 g / L, the viscosity of the immunoglobulin solution increases, causing a decrease in immunoglobulin permeability. There is.

  An inorganic salt may be added to the immunoglobulin solution as long as it does not cause denaturation or aggregation of the immunoglobulin and does not affect the permeation of a protein having a lower isoelectric point than the immunoglobulin. Inorganic salts also have effects such as prevention of immunoglobulin degradation and adsorption.

  Examples of the inorganic salt according to the present invention include sodium chloride, potassium chloride, magnesium chloride, calcium chloride, and magnesium sulfate. As the concentration, for example, the lower limit is preferably 0.001 mM or more, more preferably 0.01 mM or more, and still more preferably 0.05 mM. As an upper limit, 500 mM or less is preferable, More preferably, it is 100 mM or less, More preferably, 50 mM or less is good. In particular, when the upper limit is exceeded, the electrical properties in immunoglobulins are reduced, and there is a concern of affecting the permeability of proteins having a lower isoelectric point than immunoglobulins.

  The preservative according to the present invention is not particularly limited as long as it does not cause denaturation or aggregation of immunoglobulin and does not affect the permeation of a protein having a lower isoelectric point than immunoglobulin. For example, sodium azide Etc. The lower limit of the concentration is 0.001% by weight or more, preferably 0.005% by weight or more, and the upper limit is 1% by weight or less, preferably 0.5% by weight or less.

  In the present invention, the index of fractionation performance when separating an immunoglobulin monomer from an immunoglobulin solution containing an aggregate such as an immunoglobulin monomer and a dimer and a protein having a lower isoelectric point than the immunoglobulin is: The following can be considered.

When using an immunoglobulin monomer as a pharmaceutical product, it is better to remove as much as possible impurity biological components that may cause side effects. If the content of the immunoglobulin dimer is 1% or less, it is considered to be highly safe as a pharmaceutical product. Therefore, the transmittance of the immunoglobulin dimer and the transmittance of the immunoglobulin dimer that can achieve this are considered. Ratio (Immunoglobulin Dimer Permeability / Immunoglobulin Monomer Permeability = Permeability Ratio (D / M)) is preferably 0.20 or less, preferably 0.15 or less, more preferably 0. .10 or less is good. Further, when the immunoglobulin dimer contained in the immunoglobulin original solution is small, in order to reduce the immunoglobulin dimer content in the permeate to 1% or less, the transmittance of the immunoglobulin monomer and the immunoglobulin The transmittance ratio of the dimer may be 0.30 or less.
Similarly, it is desired to reduce as much as possible a protein having a lower isoelectric point than an immunoglobulin. Although it depends on the initial content, as an indicator, the ratio of the permeability of the immunoglobulin monomer to the permeability of the protein (the transmittance of the protein / the transmittance of the immunoglobulin monomer = the transmittance ratio ( P / M)) can be used, and similarly, the lower one is preferable, and depending on the protein, it should be 0.20 or less, preferably 0.15 or less.

  On the other hand, it is preferable that immunoglobulin monomer can be recovered as much as possible. Therefore, the transmittance of the immunoglobulin monomer according to the present invention is preferably 80% or more, preferably 85% or more, more preferably 90% or more.

  In the purification of monoclonal antibodies, the affinity chromatography purification step is an essential step. In particular, affinity chromatography using protein A or the like as a ligand is used. The separation principle is based on the affinity of immunoglobulin and other biological components for the ligand. Specifically, biological components including immunoglobulins, contaminating proteins, sugar chains, and nucleic acids are passed through affinity chromatography to specifically bind only to the immobilized immunoglobulin. The specifically bound immunoglobulin is removed from the immobilized ligand using a low pH, high pH, high salt, competitive ligand, etc., and recovered to obtain a purified immunoglobulin. However, it should be noted in the affinity chromatography purification step that it is necessary to contact low pH and high salt in order to elute the immunoglobulin from the ligand. There have been problems such as formation of immunoglobulin aggregates as a by-product and formation of aggregates with immunoglobulins by elution of a ligand (for example, protein A). Accordingly, immunoglobulins can be sufficiently purified by carrying out the separation method according to the present invention after performing affinity chromatography purification using protein A as a ligand.

  Protein A according to the present invention includes protein A recovered from natural sources, synthetically produced protein A (eg, by peptide synthesis or by recombinant techniques), and CH2 / CH3 regions (eg, And variants thereof that retain the ability to bind to proteins having the Fc region). Protein A can be purchased commercially from Repligen, Pharmacia and Fermate. Protein A is generally immobilized as a ligand on a solid support material. Furthermore, protein A column refers to an affinity chromatography resin or column containing a chromatographic solid support matrix to which protein A is covalently bonded. Examples of affinity chromatography ligands other than protein A include protein L and protein G. These protein L and protein G are also those recovered from natural sources, synthetically produced (eg, by peptide synthesis or recombinant techniques), and CH2 / CH3 regions (eg, Fc And variants thereof that retain the ability to bind to a protein having a region. Furthermore, an affinity chromatography purification method using an affinity chromatography resin or column prepared by combining protein A, protein L and protein G is also included.

  The ultrafiltration membrane module in the present invention is, for example, a flat membrane or hollow fiber membrane accommodated in a casing, and at least one liquid inlet into which an immunoglobulin solution is poured into the casing. Is provided with at least one liquid outlet for deriving. The casing used for the module is assembled from one or more casing parts. The material of the casing component can be selected as necessary, such as metal, glass, thermoplastic resin, thermosetting resin, and the like. A suitable material is a thermoplastic resin material having transparency in which the inside can be observed. Specifically, polymethyl methacrylate, polystyrene, hard vinyl chloride resin, polyethylene terephthalate, polypropylene, polystyrene butadiene copolymer, polycarbonate. And polymethyl methacrylate. Particularly preferred are transparent amorphous resins, such as polystyrene butadiene copolymer, polycarbonate, and polymethyl methacrylate.

  The method of manufacturing the casing part used when assembling the casing used in the module according to the present invention is not limited as long as it can be molded. For example, welding, press molding, injection molding, reaction injection molding, super Examples include sonic pressure bonding, plasma fusion, and adhesion using an adhesive. These may be used alone or in combination of two or more. A particularly preferable method for manufacturing a casing component is a method of sealing with an injection molded product using a thermoplastic resin having transparency as a material and an appropriate adhesive.

  Casings and / or casing parts used in the modules according to the invention are surfaces which are in contact with and / or not in contact with the liquid to be separated during molding and / or after molding and / or during assembly and / or after assembly. Surface processing can be performed. There are various methods for surface treatment. For example, when hydrophilizing, hydrophilic polymer coating or surface oxidation by plasma treatment in air, etc., when hydrophobizing, water repellent and / or mold release When the agent is applied or oxygen permeation is reduced, various inorganic coats including a silicon oxide film can be applied by vapor deposition or the like. Adhesive control at the interface of the same and / or different materials is facilitated during module assembly by performing hydrophilic treatment on the casing and / or casing material, and various types of temporary use during assembly by performing hydrophobic treatment The peelability from the protective film or the like can be improved.

  The structure of the module according to the present invention differs depending on the shape of the membrane used, for example, hollow fiber or flat membrane, but the hollow fiber or flat membrane is properly contained in the casing and the immunoglobulin solution to be separated does not mix. Any structure is acceptable. Moreover, a metal mesh, a nonwoven fabric, etc. can be combined as a film | membrane holding material, and it can accommodate in a casing, and can also be modularized.

  The separation method according to the present invention is a method of separation by size fractionation and control of solution state, and purification of synthetic pharmaceuticals and various beverages such as sake, beer, wine, happoshu, tea, oolong tea, vegetable juice, fruit juice, etc. It can also be used for purposes such as purification, separation of fine particles from chemicals and treated water, separation for oil / water separation and separation of liquid and gas for the purpose of purifying separation water and sewage.

  The present invention will now be described by way of examples and comparative examples, but is not limited thereto.

[Production example of polysulfone polymer membrane]
<Method for producing hollow fiber membrane (PSf-1)>
1,650 g of N, N-dimethylacetamide (manufactured by Wako Pure Chemical Industries, Ltd., hereinafter abbreviated as DMAc), 280 g of polysulfone (P1700, UCC, abbreviated as PSf) and 110 g of polyvinylpyrrolidone (K- 90, manufactured by BASF, hereinafter abbreviated as PVP), and poured into a 5,000 × 10 ⁻⁶ m ³ reactor for a membrane stock solution. The pressure reduction and nitrogen substitution were repeated 5 times while stirring the reactor. Thereafter, the temperature of the solution in the reactor was raised to 60 ° C. to obtain a uniform PSf DMAc solution. After confirming uniform dissolution, stirring was stopped at this stage, and degassing was performed under reduced pressure. Thereafter, the pressure was returned to the same pressure as the atmospheric pressure, and a spinning membrane stock solution maintained at 60 ° C. was obtained. 550 g of DMAc was mixed with 450 g of pure water and added to a 3,000 × 10 ⁻⁶ m ³ reactor for the internal coagulation liquid. Depressurization and nitrogen substitution were repeated 5 times to obtain an internal coagulating liquid. The internal coagulating liquid and the membrane stock solution were passed through a double spinning nozzle (inner diameter 100 μm, slit width 50 μm, outer diameter 300 μm) maintained at 60 ° C. Each flow rate was appropriately adjusted according to the winding speed during spinning. The obtained hollow fiber membrane was introduced into an external coagulation liquid (pure water) in a coagulation tank maintained at 60 ° C. with an idle running distance of 0.6 m, and after coagulation was completed, it was wound up by a winding device. It was. The winding speed was 2,400 m / hour to 4,800 m / hour. Then, the obtained hollow fiber membrane was repeatedly immersed and washed using pure water at 60 ° C., and then dried for 6 hours with a hot air dryer at 70 ° C. By this production method, a polysulfone polymer membrane having a molecular weight cut-off of 360,000, an inner diameter of 207 μm, and a film thickness of 41 μm could be produced.

<Method for producing hollow fiber membrane (PSf-2)>
By changing the composition of the internal coagulation solution (pure water / DMAc) and the polysulfone concentration in the membrane stock solution under the same conditions as the production method of PSf-1, the molecular weight cut off was 600,000, the inner diameter was 155 μm, and the film thickness was 33 μm. A polysulfone polymer membrane could be produced.

<Method for producing hollow fiber membrane (PSf-3)>
By changing the composition of the internal coagulation solution (pure water / DMAc) and the polysulfone concentration in the membrane stock solution under the same conditions as the production method of PSf-1, the molecular weight cut-off is 25,000, the inner diameter is 201 μm, the film thickness A 49 μm polysulfone polymer membrane could be produced.

<Preparation of human immunoglobulin solution>
Human immunoglobulin 5% solution (Globenin-I-Nichiyaku, Nippon Pharmaceutical Co., Ltd., isoelectric point pI7.2, abbreviated as h-IgG) was centrifuged, insoluble matter was centrifuged, and the supernatant was micronized to 0.2 μm. Filtered with a filter. In addition, as a protein having a lower isoelectric point than immunoglobulin, a bovine serum albumin 5% solution (manufactured by SIGMA-Aldrich, abbreviated as isoelectric point pI4.7, BSA) was similarly centrifuged, and the insoluble matter was centrifuged. The clear was filtered through a 0.2 μm microfilter. Using phosphate buffered saline (Nissui Co., Ltd., abbreviated as PBS), the solution was adjusted so that the h-IgG concentration was 10 g / L and the BSA concentration was 3 g / L. Using a predetermined buffer solution, diafiltration was performed with a hollow fiber membrane (PSf-3), buffer exchange was performed, and a human immunoglobulin solution was prepared. It was confirmed by absorbance meter, high performance liquid chromatography measurement, and reverse phase chromatography measurement that the concentration of each protein did not change even after diafiltration. The ratios of human immunoglobulin monomer and dimer after solution preparation were 91% and 8%, respectively.

<Production example of anti-SCF antibody (monoclonal antibody)>
(1) Preparation of immunogen SCF cDNA isolated from a cDNA library of HeLa cells that highly express SCF was incorporated into an animal cell expression vector pBCMGS-neo, which was then used as mouse fibroblast cell line Balb / 3T3 cells. The resulting transfectant was used as an immunogen.
(2) Production of hybridoma (a) Immunization The above transfectants were intraperitoneally administered to 8-week-old Balb / c mice (female) at intervals of 2 weeks. The effect of immunity was evaluated by the reactivity of peripheral blood serum collected from the tail vein of mice with the immunogen. After confirming the effect, final immunization and cell fusion were performed.
(B) Cell fusion Four days after the final immunization, the spleen cells of the immunized mouse and the mouse myeloma-derived cell line SP-2 were subjected to cell fusion according to a conventional method.
(C) Screening of anti-SCF antibody-producing hybridoma As an anti-SCF antibody-producing hybridoma screening method, an indirect antibody method using a transfectant and its parent cell line (Balb / 3T3) as an antigen was used. A hybridoma producing an antibody that binds to the transfectant and does not bind to the parent cell line (Balb / 3T3) was selected and cloned.
(D) Purification of antibody The culture supernatant of the SCF-expressing clone was concentrated by ultrafiltration, and then mixed with an equal amount of a binding buffer (BioRad Protein MAPS buffer). Protein A-Sepharose CL-4B (manufactured by Pharmacia) was equilibrated with a binding buffer, and the mixture was allowed to flow through the column to bind the antibody. Then, the column was washed with the binding buffer. An antibody fraction was obtained by flowing 0.2 M Glycine-HCl buffer (pH 3.0) through the column and elution. Next, purification was carried out in the order of DEAE-Sepharose FF (manufactured by GE Healthcare), Phenyl-Sepharose HP (manufactured by GE Healthcare), and Sephadex-G75 (manufactured by GE Healthcare), followed by anti-SCF antibody (isoelectric) Point pI 7.3) was isolated.

Next, an anti-SCF antibody is performed, the insoluble matter is centrifuged, the supernatant is filtered through a 0.2 μm microfilter, and a 5% bovine serum albumin solution (as a protein having a lower isoelectric point than immunoglobulin) ( In the same manner, SIGMA-Aldrich's isoelectric point pI 4.7-4.9, abbreviated as BSA) was centrifuged, and the supernatant was filtered through a 0.2 μm microfilter. Using phosphate buffered saline (Nissui Corp., abbreviated as PBS), the solution was adjusted so that the anti-SCF antibody concentration was 5 g / L and the BSA concentration was 1.5 g / L. Using a predetermined buffer solution, diafiltration was performed with a hollow fiber membrane (PSf-3), buffer exchange was performed, and an immunoglobulin solution was prepared. It was confirmed by absorbance meter, high performance liquid chromatography measurement, and reverse phase chromatography measurement that the concentration of each protein did not change even after diafiltration. The ratios of immunoglobulin monomer and dimer after solution preparation were 96% and 4%, respectively.

<Measurement of molecular weight cutoff>
Using each ultrafiltration membrane, 1 wt% bovine albumin (Sigma-Aldrich, molecular weight 60,000) and bovine γ-globulin (Invitrogen, molecular weight 150,000), ferritin (Sigma-Aldrich, molecular weight 45) Was filtered at a constant pressure dead end of 0.010 MPa. The amount of albumin, γ-globulin, and ferritin in the immunoglobulin permeate that permeated within 5 minutes from the start of filtration was measured, and the capture rate of each protein captured on the membrane was calculated. A calibration curve between the molecular weight and the capture rate of each protein was prepared, the molecular weight at the capture rate of 90% was determined from the calibration curve, and the value was determined as the fractional molecular weight.

<Calculation method of transmittance and transmittance ratio>
Examples of the method for measuring the transmittance and the transmittance ratio include a method of calculating from results such as high performance liquid chromatography, nuclear magnetic resonance, mass spectrometry, infrared spectroscopy, etc. However, the present invention is not limited to these. In the present invention, the transmittance and the transmittance ratio were calculated by the following method. The immunoglobulin concentration was calculated by measuring the absorbance with an absorptiometer using a wavelength of 280 nm. First, the concentration of BSA was measured using reverse phase chromatography (LC-10 manufactured by Shimadzu Corporation, columns manufactured by Shimadzu Corporation: Imtakt, PretoFT-C18, gradient of distilled water / acetonitrile). It was determined from the absorption peak area ratio. The weight ratio of immunoglobulin monomer to dimer was measured by high performance liquid chromatography (HPLC) (two columns G3000SWXL manufactured by Tosoh Corporation, SC8020 system manufactured by Tosoh Corporation, manufactured by Tosoh Corporation). UV8020 detector) and obtained from the absorption peak area ratio at a wavelength of 280 nm. When the peak of BSA and immunoglobulin monomer overlap, the BSA peak area detected by high performance liquid chromatography is calculated from the BSA concentration calculated by the reverse phase chromatography method and easily subtracted by immunoglobulin 1 The peak area of the polymer can be calculated.

(Transmittance calculation method)
First, the total throughput of immunoglobulin permeated through the membrane can be calculated by the following formulas (29) to (31).
W = W (Mo) + W (Ag) (29)
W (Mo)
= V1 * S1 / Sg * G1 (Mo) -V2 * S2 / Sg * G2 (Mo) (30)
W (Ag)
= V1 * S1 / Sg * G1 (Ag) -V2 * S2 / Sg * G2 (Ag) (31)

W: Amount of immunoglobulin processed through the membrane (g)
W (Mo): treatment amount of immunoglobulin monomer permeating through the membrane (g)
W (Ag): throughput of immunoglobulin dimer permeated through the membrane (g)
Sg: HPLC peak area of all immunoglobulins at 1 g / L (mV · sec)
V1: Volume of the original immunoglobulin solution before filtration (L)
S1: HPLC peak area (mV · sec) of all immunoglobulins in the original solution before filtration
G1 (Mo): Monomer content (%) in the immunoglobulin solution of the original solution before filtration
G1 (Ag): Dimer content (%) in the immunoglobulin solution of the original solution before filtration
V2: Volume of the original immunoglobulin solution after filtration (L)
S2: HPLC peak area (mV · sec) of all immunoglobulins in the original solution after filtration
G2 (Mo): Monomer content (%) in the immunoglobulin solution of the original solution after filtration
G2 (Ag): Dimer content (%) in the immunoglobulin solution of the original solution after filtration

However, when the volume of the original immunoglobulin solution before filtration is large and the treatment amount is small, the composition ratio of the monomer and the dimer in the immunoglobulin solution hardly changes. Therefore, as an approximate value, the following formula (32) The processing amount may be calculated from (33).
W (Mo)
= (V1 * S1-V2 * S2) / Sg * G1 (Mo) / 100 (32)
W (Ag)
= (V1 * S1-V2 * S2) / Sg * G1 (Ag) / 100 (33)

Next, the permeation amount P (Mo) of the immunoglobulin monomer penetrating the membrane and the permeation amount P (Ag) of the dimer were calculated by the following formulas (34) and (35).
P (Mo) = V3 × S3 / Sg × G3 (Mo) (34)
P (Ag) = V3 × S3 / Sg × G3 (Ag) (35)
P (Mo): Permeation amount of immunoglobulin monomer passing through the membrane (g)
P (Ag): Amount of immunoglobulin dimer permeating through the membrane (g)
Sg: HPLC peak area of all immunoglobulins at 1 g / L (mV · sec)
V3: Volume of immunoglobulin solution in permeate (L)
S3: HPLC peak area (mV · sec) of all immunoglobulins in the permeate
G3 (Mo): Monomer content (%) in the immunoglobulin solution of the permeate
G3 (Ag): Dimer content (%) in the immunoglobulin solution of the permeate

Further, the monomer transmission rate and the dimer transmission rate in the immunoglobulin solution were calculated by the following formulas (36) and (37), and the immunoglobulin dimer and monomer transmission ratio (Ag / Mo) Was calculated by the following formula (38).
Monomer transmittance (%) = P (Mo) / W (Mo) × 100 (36)
Dimer transmittance (%) = P (Ag) / W (Ag) × 100 (37)
Transmittance ratio (Ag / Mo) = dimer transmittance / monomer transmittance (38)

The lower the transmittance ratio (Ag / Mo), the lower the transmittance of the dimer, the higher the transmittance of the monomer, and the higher the fractionation performance of the monomer and the dimer. Yes.

In order to compare the separation performance of BSA and immunoglobulin monomer, the transmittance ratio (B / Mo) between BSA and immunoglobulin monomer was calculated by the following formula (39).
Transmittance ratio (B / Mo)
= (S4 * S1 * G1 (Mo)) / (S5 * S3 * G3 (Mo)) (39)

S4: Peak area (mV · sec) of reverse phase chromatography of BSA in permeate
S5: Peak area (mV · sec) of BSA reverse phase chromatography of the original solution before filtration
S1: HPLC peak area (mV · sec) of all immunoglobulins in the original solution before filtration
G1 (Mo): Monomer content (%) in the immunoglobulin solution of the original solution before filtration
S3: HPLC peak area (mV · sec) of all immunoglobulins in the permeate
G3 (Mo): Monomer content (%) in the immunoglobulin solution of the permeate

The lower the permeability ratio (B / Mo), the lower the permeability of BSA, the higher the permeability of immunoglobulin monomer, and the higher the fractionation performance of immunoglobulin monomer and BSA. Yes.

[Example 1]
The number was taken out so that the total cross-sectional area of the hollow portion of the hollow fiber membrane (PSf-1) was 0.02 m ^2, and a yarn bundle for evaluation of immunoglobulin monomer separation and purification was produced. Next, 10 g / L human immunoglobulin (h-IgG), 3 g / L bovine serum albumin (BSA), pH 8.9, 1.5 mM tris (hydroxymethyl) aminomethane-hydrochloric acid (Tris) buffer solution were added. An immunoglobulin solution having a pH of 8.9 was prepared. The yarn bundle and 100 mL of the human immunoglobulin solution are connected to a crossflow filtration device as shown in FIG. 3 so that the linear velocity in the hollow fiber membrane is 10 cm / second and the outlet pressure of the hollow fiber membrane is 0.010 MPa. The liquid feed pump 2 (3) was rotated and adjusted with the adjustment valve (7). Since the protein concentration in the immunoglobulin solution that undergoes cross-flow filtration is increased, 1.5 mM tris (hydroxymethyl) aminomethane having a pH of 8.9 in the diluent tank (1) so that the concentration is always constant. Hydrochloric acid (Tris) buffer is added as a diluent to the immunoglobulin source solution tank (4). Crossflow filtration was performed at 25 ° C. until no solution was present, and the permeate was collected. The permeate was analyzed by reverse phase chromatography measurement and high performance liquid chromatography measurement, and the transmittance of immunoglobulin monomer and dimer, the transmittance of BSA, and the transmittance ratio of each were calculated.
As a result (Table 1), h-IgG monomer and dimer transmittances were 92.5% and 9.3%, respectively, and the transmittance of BSA was 11.1%. The transmittance ratio between h-IgG dimer and monomer was 0.10, and the transmittance ratio between BSA and h-IgG monomer was 0.12. This result shows that the h-IgG dimer can be recovered with high transmittance, and the h-IgG dimer and BSA can be removed.

[Example 2]
As an immunoglobulin solution, 5 g / L SCF monoclonal antibody (SCF) and 1.5 g / L bovine serum albumin (BSA), pH 8.9, 1.5 mM tris (hydroxymethyl) aminomethane-hydrochloric acid (Tris) buffer Using the solution, cross flow filtration was performed in the same manner as in Example 1 except that an immunoglobulin solution having a pH of 8.9 was prepared.
As a result (Table 1), the SCF monomer and dimer transmittances were 91.4% and 13.1%, respectively, and the transmittance of BSA was 10.8%. The transmittance ratio between the SCF dimer and the monomer was 0.14, and the transmittance ratio between the BSA and the SCF monomer was 0.12. This result shows that the SCF monomer can be recovered with high permeability, and the h-IgG dimer and BSA can be removed.

[Comparative Example 1]
Cross flow filtration was performed in the same manner as in Example 1 except that the hollow fiber membrane (PSf-1) was changed to the hollow fiber membrane (PSf-2).
As a result (Table 1), the h-IgG monomer and dimer transmittances were 97.6% and 54.2%, respectively, and the transmittance of BSA was 98.0%. The transmittance ratio between the h-IgG dimer and the monomer was 0.56, and the transmittance ratio between the BSA and the h-IgG monomer was 1.00. This result shows that the h-IgG dimer and BSA cannot be separated and removed, although the h-IgG monomer can be recovered with high transmittance.

[Comparative Example 2]
In Example 1, crossflow filtration was performed by the same method as Example 1 except that the pH of the immunoglobulin solution was 6.9.
As a result (Table 1), the h-IgG monomer and dimer transmittances were 92.7% and 10.6%, respectively, and the BSA transmittance was 94.0%. The transmittance ratio between h-IgG dimer and monomer was 0.11, and the transmittance ratio between BSA and h-IgG monomer was 1.01. This result shows that the h-IgG monomer can be recovered with high transmittance and the h-IgG dimer can be removed, but BSA cannot be separated and removed.

[Comparative Example 3]
The buffer flow was not changed during the preparation of the immunoglobulin solution, and the cross flow was performed in the same manner as in Example 1 except that the buffer solution of pH 139 mM PBS was used as the pH 7.0 immunoglobulin solution. Filtration was performed.
As a result (Table 1), the h-IgG monomer and dimer transmittances were 94.5% and 9.2%, respectively, and the transmittance of BSA was 97.5%. The transmittance ratio between h-IgG dimer and monomer was 0.10, and the transmittance ratio between BSA and h-IgG monomer was 1.03. This result shows that the h-IgG monomer can be recovered with high transmittance and the h-IgG dimer can be removed, but BSA cannot be separated and removed.

The separation method according to the present invention can be suitably used in the field of separation and purification of biopharmaceuticals such as immunoglobulins.

It is a figure which illustrates the crossflow filtration apparatus of this invention. It is a figure which illustrates the crossflow filtration apparatus of this invention. It is a figure which illustrates the cross flow filtration method of the present invention.

Explanation of symbols

1 Diluent tank 2 Liquid feed pump 1
3 Liquid feed pump 2
4 Immunoglobulin source solution tank 5 Pressure gauge 1
6 Pressure gauge 2
7 Adjustment valve 8 Ultrafiltration membrane module 9 Immunoglobulin permeate tank 10 Flow meter 11 Concentration controller 12 Pressure / flow rate controller 13 UV flow cell

Claims (15)

  Using an ultrafiltration membrane having a molecular weight cut off of 100,000 or more and less than 500,000, an immunoglobulin solution comprising at least an immunoglobulin monomer, an aggregate thereof, and a protein having a lower isoelectric point than immunoglobulin Separation characterized in that the immunoglobulin monomer is separated by cross-flow filtration of an immunoglobulin solution having an immunoglobulin concentration of 1-150 g / L and a pH equal to or higher than the isoelectric point of the immunoglobulin. Method.   The separation method according to claim 1, wherein the aggregate contains at least an immunoglobulin dimer.   3. The separation method according to claim 1, wherein tris (hydroxymethyl) aminomethane is used as a buffer for adjusting the pH of the immunoglobulin solution.   The separation method according to any one of claims 1 to 3, wherein the concentration of the buffer used in the immunoglobulin solution is 0.01 mM to 500 mM.   The separation method according to any one of claims 1 to 4, wherein the pH of the immunoglobulin solution is from the isoelectric point of the immunoglobulin to 11 or less.   The separation method according to any one of claims 1 to 5, wherein the protein having a lower isoelectric point than the immunoglobulin is albumin.   The separation method according to claim 1, wherein the immunoglobulin concentration is 5 to 100 g / L.   The separation method according to any one of claims 1 to 7, wherein the cross-flow filtration is performed while maintaining the immunoglobulin concentration substantially constant.   The separation method according to any one of claims 1 to 8, wherein the immunoglobulin is a monoclonal antibody.   The separation method according to claim 1, wherein the ultrafiltration membrane is a polysulfone polymer membrane. The separation method according to claim 10, wherein the polysulfone polymer is at least one or a mixture of two or more polysulfone polymers represented by the following formulas (1) to (3).
[Chemical 1]

[Chemical 2]

[Chemical 3]

  The separation method according to claim 10 or 11, wherein the polysulfone polymer is a polysulfone polymer hydrophilized with polyvinylpyrrolidone.   The separation method according to claim 1, wherein the ultrafiltration membrane is a hollow fiber membrane. The separation method according to any one of claims 1 to 13, which is carried out using an apparatus comprising one or more means selected from the following (a) to (d).
(B) Means capable of monitoring the concentration of the original immunoglobulin solution (b) Means capable of controlling the concentration of the immunoglobulin original solution (c) Means capable of controlling the linear velocity of the immunoglobulin original solution (d) Filtration pressure of the ultrafiltration membrane Means to control An apparatus comprising one or more means of the module used in the separation method according to any one of claims 1 to 14 and means comprising the following (a) to (d).
(B) Means capable of monitoring the concentration of the original immunoglobulin solution (b) Means capable of controlling the concentration of the immunoglobulin original solution (c) Means capable of controlling the linear velocity of the immunoglobulin original solution (d) Filtration pressure of the ultrafiltration membrane Means to control

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