JPWO2013176140A1 - Method and apparatus for producing high concentration protein solution - Google Patents

Abstract

Concentrate dilute protein solutions such as ascites to obtain a concentrated protein solution, without reducing the concentration efficiency due to clogging, and obtaining a concentrated protein solution with a high protein concentration without additional processing time such as an additional concentration step An object of the present invention is to provide a method for producing a highly concentrated protein solution. The present invention relates to a polysulfone-based hollow fiber membrane type in which a low-concentration protein solution is passed through a circuit from a storage container in which the low-concentration protein solution is stored, and an ultrafiltration performance is 85 mL to 150 mL / min / 200 mmHg and a hydrophilic polymer is added. The first step of allowing the ascites concentration filter to pass through and sending the filtrate from the filtration side outlet of the filter, and sending the high concentration protein solution from the filter outlet, and the high concentration protein solution sent from the filter outlet A first step of passing the low concentration protein solution through the ascites concentration filter at a first flow rate, and a second step of recovering the low concentration protein solution. When a predetermined amount or more is delivered from the total amount, the low-concentration protein solution is passed through the ascites concentration filter at a second flow rate that is faster than the first flow rate. Comprising a second step, the a process for producing a high concentration protein solution. [Selection] Figure 1

Description

  The present invention relates to a method and an apparatus for producing a high concentration protein solution.

Conventionally, for patients who have a tendency to accumulate ascites or pleural effusion (hereinafter collectively referred to as ascites) such as cirrhosis, the protein in the ascites is used to increase the patient's blood protein concentration. Ascites drained into the ascites is filtered and concentrated with two types of filters using hollow fiber membranes, etc. to obtain a concentrated protein solution, and this is instilled into the patient ascites filtered concentrated re-injection method (for example, (See Patent Documents 1 and 2). The first of the two types of filters is a filter for removing cell components such as cancer cells and blood cell components contained in ascites, and does not allow cell components to pass through, but allows solute components such as moisture and protein to pass through. A membrane having such a pore size is used. On the other hand, the other filter is a concentration filter for dehydrating ascites, which is a dilute protein concentration, and concentrating the protein. A membrane that hardly passes protein components and allows moisture, electrolytes, etc. to pass through is used. In general, from the viewpoint of convenience, a method of concentrating ascites obtained by filtering cell components with a filter with a concentrator is used, and a device that performs these continuously is used.
On the other hand, as means for suppressing protein leakage, in the field of hemodiafiltration, there is a method of controlling the membrane TMP by using a device to control the flow rate of the membrane and realizing optimal diafiltration. . (For example, Patent Document 3).

JP 2009-297242 A Utility model registration No. 2543466 Japanese Patent Laid-Open No. 2001-112863

  The protein solution to be administered to a patient also has a purpose of increasing the blood protein concentration of the patient after administration, and therefore needs to be at a somewhat high concentration. For this purpose, it needs to be concentrated to a high concentration ratio. When the protein concentration of the ascites to be processed becomes high, the concentration filter may be clogged with protein and the concentration efficiency may be reduced, so that concentration to the target concentration ratio may not be achieved. In such a case, it is necessary to go through a very complicated process such as collecting the protein solution in an insufficiently concentrated state and then concentrating it additionally. In some cases, it may be necessary to replace the concentration filter with a new one. This is a burden on the operator, and there is also an economic disadvantage. In addition, for the patient, additional processing time is required, so that the binding time until the protein solution is administered becomes longer, or when the protein is recovered during additional concentration, the protein is lost and the dosage is reduced. There is a disadvantage of doing.

  On the other hand, patients with ascites can be broadly divided into patients with hepatic ascites that accumulates due to diseases such as cirrhosis and those with cancer ascites that retains cancers such as stomach cancer, ovarian cancer, and colon cancer. In the past, this treatment was mostly performed mainly for patients with hepatic ascites, but in recent years, the therapeutic effect of implementing this treatment for patients with cancerous ascites has been recognized. Opportunities for patients with ascites are increasing. However, since cancerous ascites generally tends to have a higher protein concentration than hepatic ascites, the additional concentration step as described above is frequently required.

  Further, Patent Document 1 discloses a method in which a negative pressure generator is installed downstream of a concentration filter to suppress accumulation of deposits in a method of concentration using a suction device provided on the waste liquid side of the concentration filter. However, also in this method, it is necessary to take action by an operator in order to open the pressure release line provided in the middle of the suction device and release the suction pressure applied to the concentrator.

  Furthermore, when the ascites filtration concentration process is performed, if the ascites inflow rate is low, the processing time becomes long, and the restraint time of the patient is extended to increase the burden. On the other hand, when the inflow rate of ascites is increased, useful proteins may leak to the filtrate side of the ascites concentration filter.

  The present invention is a method for concentrating a dilute protein solution such as ascites to obtain a concentrated protein solution in response to the problems of the conventional method described above, without causing a reduction in processing speed and without burden on the operator such as an additional concentration step. It is an object of the present invention to provide a method and an apparatus for producing a high concentration protein solution capable of obtaining a concentrated protein solution having a high protein concentration.

As a result of intensive studies on the above problems, the present inventors have found that the clogging is related to the ultrafiltration performance of the concentration filter, and by using a concentration filter having a specific ultrafiltration performance, the decrease in the concentration rate is suppressed. In addition, by controlling the flow rate through the concentration filter in at least two steps, it was found that the protein can be concentrated to a high magnification in a short time without losing the protein, and the present invention has been completed.
In addition, it is desirable to control the flow rate when the sieving coefficient of protein or the weight of the original ascites falls below a certain level. It was difficult to measure. On the other hand, as a factor to change the numerical value of the sieving coefficient, not only the permeation performance of the filter itself but also the influence of the membrane area and the amount of treated ascites was experimentally found, and the control of the flow rate was simplified. The present invention has been completed with parameters to do so.

  That is, the present invention relates to the following [1] to [16].

[1]
Aspirate the low-concentration protein solution through a circuit from a storage container in which the low-concentration protein solution is stored, using a polysulfone-based hollow fiber membrane type ascites with an ultrafiltration performance of 85 mL to 150 mL / min / 200 mmHg and a hydrophilic polymer. A first step of letting the solution pass through and letting out the filtrate from the filtration side outlet of the filter and sending out the high concentration protein solution from the outlet of the filter;
A second step of collecting the high concentration protein solution delivered from the outlet of the filter in a collection container;
Including
The first step includes
A first step of passing the low concentration protein solution through the ascites concentration filter at a first flow rate;
A second step of passing the low-concentration protein solution through the ascites concentration filter at a second flow rate faster than the first flow rate when a predetermined amount or more is sent from the total amount of the low-concentration protein solution;
A method for producing a high concentration protein solution, comprising:

[2]
The membrane area of the filter is A, the weight of the low-concentration protein solution treated in the first step is V1, the protein concentration of the low-concentration protein solution is C, the flow rate of the first step is Qb1, and the first step The method according to [1], wherein when the filtration flow rate in step 1 is Qf1 and the ultrafiltration performance of the filter is F, the process is switched to the second step at a timing satisfying the following (1).
103.7 ≦ −37log (A / V1) + log (Qb1 / V1) + 57log (F) −log (1 / C) −log (Qb1 / Qf1) ≦ 112.6 (1)

[3]
The method of [1] or [2], wherein the switching from the first flow rate to the second flow rate is performed when at least 1/4 of the total amount of the low concentration protein solution is fed.

[4]
The method according to any one of [1] to [3], wherein the first flow rate is 70 mL / min or less and the second flow rate is 120 mL / min or less.

[5]
The method according to any one of [1] to [4], wherein the first flow rate is 50 mL / min or less and the second flow rate is 70 mL / min or less.

[6]
The total value of the first flow rate and the second flow rate is 100 mL / min or more, and the flow rate difference between the first flow rate and the second flow rate is at least 20 mL / min or more [1] to [ 5].

[7]
The method according to any one of [1] to [6], wherein in the first step, the protein concentration of the low concentration protein solution is 5 g / dL or less.

[8]
The method according to any one of [1] to [6], wherein in the first step, the protein concentration of the low concentration protein solution is 3 g / dL or less.

[9]
In the first step, when the protein sieving coefficient in the high-concentration protein solution delivered from the filtration-side outlet of the ascites concentration filter is less than or equal to a predetermined value, the second step is started [1] Any method of [8].

[10]
In the first step, when the sieving coefficient of the protein in the high concentration protein solution delivered from the filtration side outlet of the ascites concentration filter is at least 0.03 or less, the second step is started. [1] to [9].

[11]
The method according to any one of [1] to [10], wherein in the first step, the protein concentration in the filtrate sent from the filtration-side outlet of the ascites concentration filter is 100 mg / dL or less.

[12]
The method according to any one of [1] to [11], wherein in the first step, the protein concentration of the high concentration protein solution delivered from the outlet of the ascites concentration filter is 7 g / dL or more.

[13]
The method according to any one of [1] to [12], wherein the circuit includes an ascites filter.

[14]
In the first step, the first step of feeding the ascites concentration filter to the ascites concentration filter at a first flow rate, the protein concentration in the solution at the inlet and the outlet of the ascites concentration filter, and monitoring the protein The method according to any one of [1] to [13], wherein the second step is controlled when the coefficient has decreased to at least 0.03 or less.

[15]
In the first step, the first step of sending the liquid to the ascites concentration filter at a first flow rate, and monitoring the weight of the low concentration protein solution, a predetermined amount or more from the total amount of the low concentration protein solution is The method according to any one of [1] to [13], wherein control is performed so that the second step is performed when the liquid is fed.

[16]
Aspirate the low-concentration protein solution through a circuit from a storage container in which the low-concentration protein solution is stored, using a polysulfone-based hollow fiber membrane type ascites with an ultrafiltration performance of 85 mL to 150 mL / min / 200 mmHg and a hydrophilic polymer. A part having a first step of letting the solution pass through and letting out the filtrate from the filtration side outlet of the filter and sending out the high concentration protein solution from the outlet of the filter;
A portion having a second step of collecting the high-concentration protein solution delivered from the outlet of the filter in a collection container;
Including
The part having the first step comprises
A portion having a first step of passing the low concentration protein solution through the ascites concentration filter at a first flow rate;
A second step of passing the low-concentration protein solution through the ascites concentration filter at a second flow rate faster than the first flow rate when a predetermined amount or more is delivered from the total amount of the low-concentration protein solution. And
Including
The membrane area of the filter is A, the weight of the low-concentration protein solution treated in the first step is V1, the protein concentration of the low-concentration protein solution is C, the flow rate of the first step is Qb1, and the first step An apparatus for producing a high-concentration protein solution that switches to the second step at a timing that satisfies the following (1), where Qf1 is the filtration flow rate of step 1 and F is the ultrafiltration performance of the filter.
103.7 ≦ −37log (A / V1) + log (Qb1 / V1) + 57log (F) −log (1 / C) −log (Qb1 / Qf1) ≦ 112.6 (1)

  According to the present invention, in a method for concentrating a dilute protein solution such as ascites and obtaining a concentrated protein solution, the processing speed is not reduced due to clogging, and a high protein concentration is obtained without burden on the operator such as an additional concentration step. It becomes possible to provide a manufacturing method and a manufacturing apparatus of a high concentration protein solution that can obtain a concentrated protein solution.

It is a figure which shows 1st Embodiment of an ascites filtration concentration apparatus. It is a figure which shows 2nd Embodiment of an ascites filtration concentration apparatus. It is a figure which shows 3rd Embodiment of an ascites filtration concentration apparatus. It is a figure which shows the ascites filtration concentration apparatus provided with the refractometer. It is a figure which shows the ascites filtration concentration apparatus provided with the control apparatus (for weight monitoring). It is a figure which shows the test apparatus of the ascites concentration performance based on the filter for concentration.

  Hereinafter, a mode for carrying out the present invention (hereinafter referred to as “the present embodiment”) will be described in detail. In addition, this invention is not limited to the following embodiment, It can implement by changing variously within the range of the summary.

(First embodiment)
As a first embodiment of the present invention, an example of the configuration of the ascites filtration concentration device 100 shown in FIG. 1 and a method for obtaining a concentrated protein solution (high concentration protein solution) by filtering and concentrating ascites (protein solution) using the device 100. Indicates.

  First, in this embodiment, a low concentration protein solution that is ascites collected in advance from a patient is stored in the storage container 1. In general, ascites is a low concentration protein solution with a protein concentration of 5 g / dL or less. When the protein concentration exceeds 5 g / dL, the time required to form a dense blood layer in the hollow fiber due to the fouling phenomenon is short, so even if the flow rate is relatively high from the beginning, the protein leaks to the filtrate side. Does not occur much, and as a result, the method of the present invention does not need to be applied. Similarly, when the protein concentration exceeds 3 g / dL, although not as much as when it exceeds 5 g / dL, the same phenomenon occurs in a relatively short time, so there is a case where it is not always necessary to apply the method of the present invention. is there. Therefore, the low concentration protein solution in the present embodiment preferably has a protein concentration of 5 g / dL or less, more preferably 3 g / dL or less.

  The outlet 1 b of the storage container 1 is connected to the first flow path 31 and is connected to the inlet 3 a of the filter 3 for filtration via the first flow path 31. Ascites fluid in the storage container 1 is supplied to the filter 3 for filtration via the first flow path 31. In the filter 3 for filtration, since the cell component cannot pass through the filtration membrane, the ascites after filtration including the protein from which the cell component has been removed is discharged from the filtrate outlet 3c of the filter 3 for filtration. A third flow path 33 is connected to the outlet 3 b of the filter 3 for filtration, and a part of ascites containing cell components is discharged out of the device 100 through the third flow path 33. The filtrate outlet 3 c is connected to the ascites inlet 4 a of the concentration filter 4 via the second flow path 32, and the post-filtered ascites discharged from the filtrate outlet 3 c is concentrated via the second flow path 32. Introduced into the filter 4.

  In the concentration filter 4, the ascites water after filtration is separated and discharged from the apparatus 100 through the fifth flow path 35 connected to the filtrate outlet 4 c. Here, the liquid discharged from the filtrate outlet 4c is preferably a low concentration protein solution having a protein concentration of 100 mg / dL or less. The pore diameter of the concentration filter 4 is smaller than the pore diameter of the filtration filter 3. Since the protein contained in the ascites after filtration does not pass through the filtration membrane and is retained in the concentration filter 4, the protein concentration in the ascites after filtration is increased by the drainage of the water, and the concentration filter 4 is concentrated as a concentrated protein solution. From the concentrated liquid outlet 4b. The concentrate outlet 4b of the concentration filter 4 is connected to the inlet 2a of the collection container 2 via the fourth channel 34, and the concentrated protein solution discharged from the concentrate outlet 4b passes through the fourth channel 34. Through the collection container 2. The concentrated protein solution recovered here is a high-concentration protein solution having a protein concentration higher than at least ascites in the storage container 1, and preferably has a protein concentration of 7 g / dL or more.

  The storage container 1 and the recovery container 2 may be anything as long as they can store a liquid, but usually a polyvinyl chloride bag is used from the viewpoint of handleability. The size of the container is determined by the amount of ascites stored.

  The filter 3 for filtration is not particularly limited as long as it can separate cell components and moisture, and solute components such as electrolytes and proteins. The structure, shape, and dimensions of the filter are not limited as long as it includes an ascites inflow port that can be connected to the flow path connected to the storage container 1 or the concentration filter 4 and a filtered ascites outlet. Further, the hollow fiber used in the filter 3 for filtration is not particularly limited, and for reasons of easy pore size control during film formation and excellent chemical stability, polyolefins such as polyethylene, polysulfones, regenerated celluloses, Polyvinyl alcohol is preferred. These exemplified hollow fiber materials may contain other materials or may be chemically modified. Usually, a hollow fiber membrane filter having a pore size of 0.2 μm or less and a protein permeability of 80% or more is used.

  In FIG. 1, the filter 3 for filtration is used by the internal pressure filtration method, and ascites filtration is performed. However, if the cell component can be separated, the external pressure filtration method can be used. It can be sealed and filtered by a dead end method, or the third flow path 33 can be opened to be a cross flow method.

  As long as each flow path 31-35 which comprises the apparatus of 1st Embodiment can also connect with the storage container 1, the collection container 2, the filter 3 for filtration, and the filter 4 for concentration, there is no limitation in a material, a dimension, etc. Usually, a soft tube manufactured from polyvinyl chloride or the like is used as a tube forming the flow paths 31 to 35.

Any means may be used for feeding ascites from the storage container 1 to the filter 3 for filtration. For example, as shown in FIG. 1, a pump 5 may be installed in the first flow path 31 to send liquid. As the pump 5, a roller pump or an infusion pump is generally used. Further, a liquid feed pump may be added also on the second flow path 32 and the third flow path 33.
In FIG. 1, the control unit 14 is installed in the middle of the fourth flow path 34, and the control unit 15 is installed in the middle of the fifth flow path 35. The control unit 14 adjusts the flow rate of the fourth flow path 34, and the control unit 15 adjusts the flow rate of the fifth flow path 35. The control units 14 and 15 can adjust the balance between the amount of the filtrate discharged from the concentration filter 4 and the amount of the concentrated protein solution, and adjust the concentration ratio.

  The control units 14 and 15 balance the amount of filtrate that is filtered from the concentration filter 4 and discharged from the concentrated solution outlet 4b in the ascites supplied to the concentration filter 4 and the amount of liquid that is directed to the collection container 2. Anything can be used as long as it can be controlled. For example, the control units 14 and 15 may control the flow path resistance by adjusting a flow path resistance such as a roller clamp, and apply a certain negative pressure or positive pressure to each of the flow paths 34 and 35. It may be a device that controls the flow rate, or may be a device that controls the flow rate, such as a roller pump or an infusion pump. As long as the balance between the amount of liquid filtered and discharged from the concentration filter 4 and the amount of liquid directed to the collection container can be controlled, only one of the controllers 14 and 15 may be used.

The control device 41 controls the flow rate of ascites in the first flow path 31 (the amount of liquid delivered per unit time) by controlling the driving of the pump 5. The control device 41 is, for example, a computer, and may also serve as an input terminal that allows an enforcer to input information related to desired control. In addition, the flow rate of the flow paths 34 and 35 may be controlled by the control device 41, assuming that the control units 14 and 15 are driven by a control signal from the control device 41.
The control device 41 controls the flow rate when the numerical value exceeds a certain value, for example, by monitoring the weight of the low concentration protein solution and the protein concentration.

  In the apparatus 100 shown in FIG. 1, the first flow path 31 is connected to the lower side of the filtration filter 3 and the third flow path 33 is connected to the upper side of the filtration filter 3. The effect is obtained. The second flow path 32 may be connected to any position as long as it communicates with the hollow fiber outer chamber portion of the filter 3 for filtration.

  The ultrafiltration performance of the concentration filter 4 used in this embodiment is 85 mL to 150 mL / min / 200 mmHg. If the ultrafiltration performance is less than this range, the amount of filtrate discharged during concentration is reduced, and a sufficiently concentrated protein solution cannot be obtained. Moreover, if it is 95 mL / min / 200 mmHg or more, there is little possibility of clogging and it is preferable. If it is greater than 150 mL / min / 200 mmHg, the protein leaks into the filtrate, and a sufficient protein concentration cannot be obtained, which is not suitable.

The ultrafiltration performance in the present embodiment is defined by the following test. Prepare bovine plasma with a protein concentration adjusted to 6 g / dL, and send it to a concentration filter at a constant speed of 200 mL per minute with a roller pump. At this time, the filtrate outlet of the concentrator is open. The circuit connected to the recovery liquid outlet of the concentrating filter is pressed and adjusted so that the pressure difference (hereinafter also referred to as TMP) applied to the inside and outside of the filter becomes 200 mmHg. At this time, the volume per hour of the filtrate discharged from the filtrate outlet is measured. TMP is calculated as follows.
TMP = (pressure on the filter inlet side + pressure on the filter outlet side) / 2-filtrate side pressure

  In this embodiment, a filter using a hollow fiber membrane is used from the viewpoint of concentration efficiency. The hollow fiber membrane here is not particularly limited in its shape and dimensions, and any hollow fiber membrane may be used as long as it has the above ultrafiltration performance. As the material, polysulfone is preferable because it is easy to control the pore diameter during film formation and is excellent in chemical stability. Since the polysulfone polymer is an aromatic compound, it is particularly excellent in radiation resistance, is extremely resistant to heat and chemical treatment, and is excellent in safety. Accordingly, various membrane forming conditions can be adopted and radiation sterilization can be performed, which is particularly preferable as a membrane material used in the ascites concentrator. In addition, “to system” means not only a homopolymer but also a copolymer with another monomer and a chemically modified analog.

  The polysulfone-based polymer (hereinafter sometimes referred to as PSf) as used herein is a general term for polymer compounds having a sulfone bond, and is not particularly defined. For example, the repeating unit is represented by the following formula (1 ), Formula (2), formula (3), formula (4), and polysulfone-based polymer represented by formula (5). It may be a modified polymer in which a substituent is introduced into a part of these aromatic rings. Aromatic polysulfone-based polymers represented by the formula (1), formula (2) and formula (3) are preferred from the viewpoint of industrial availability, and among them, polysulfone having a chemical structure represented by formula (1) is preferred. Particularly preferred. This bisphenol-type polysulfone resin is commercially available, for example, from Solvay Advanced Polymers under the trade name “Udel (registered trademark)”, and there are several types depending on the degree of polymerization, but there is no particular limitation.

  The polysulfone-based hollow fiber membrane in the present embodiment is made hydrophilic by a hydrophilic polymer. This is because the surface of the hollow fiber membrane becomes hydrophobic only with the polysulfone-based polymer, and the protein is easily adsorbed on such a surface, which causes a decrease in the protein recovery performance. Examples of the hydrophilic polymer include polyvinyl pyrrolidone (hereinafter sometimes referred to as PVP), polyethylene glycol, polyvinyl alcohol, polypropylene glycol, and the like. Among them, PVP is preferable from the viewpoint of hydrophilization effect and safety. There are several types of PVP depending on the molecular weight and the like. Examples of commercially available products include PVP K-15, 30, 90 (all manufactured by ISP Corporation). The molecular weight (viscosity average molecular weight) of PVP used in this embodiment is 10,000 to 2,000,000, preferably 50,000 to 1,500,000. The content of the hydrophilic polymer in the film is 3 to 20%, preferably 3 to 10% of the total amount of the polymer. When the content is 3% or less, the effect as a hydrophilizing agent is diminished, and when the content exceeds 20%, the viscosity of the film-forming stock solution is excessively increased, which is not preferable for production.

  As a method for producing a hydrophilic polysulfone hollow fiber membrane, a known dry / wet film-forming technique can be used. First, a polysulphone polymer and a hydrophilic polymer such as polovinylpyrrolidone are dissolved in a common solvent to prepare a uniform spinning solution. As such a common solvent, when the hydrophilic polymer is polyvinylpyrrolidone, for example, dimethylacetamide (hereinafter referred to as DMAC), dimethylsulfoxide, N-methyl-2-pyrrolidone, dimethylformamide, sulfolane, dioxane, etc. Or a solvent composed of a mixture of two or more of the above solvents. In order to control the pore size, additives such as water may be added to the spinning dope.

  When forming the hollow fiber membrane, a tube-in-orifice type spinneret is used, and the spinning solution from the orifice of the spinneret and the hollow internal solution from the tube are simultaneously discharged into the air. The hollow inner liquid is for coagulating the spinning dope, and water or a coagulating liquid mainly composed of water can be used. The composition of the hollow inner liquid should be determined according to the performance of the intended hollow fiber membrane, such as the ultrafiltration performance, and cannot be determined in general, but generally the solvent used for the spinning dope A mixed solution of water and water is preferably used. For example, a 0-65 wt% DMAC aqueous solution or the like is used as the hollow inner liquid. The spinning stock solution discharged from the spinneret together with the hollow inner liquid travels through the idle running portion, and is introduced and immersed in a coagulation bath mainly composed of water installed at the lower part of the spinneret to complete the coagulation. Thereafter, the solidified hollow fiber is subjected to a washing step and the like, and then wound with a wet hollow fiber membrane winder to obtain a bundle of hollow fiber membranes, and then dried. Alternatively, a hollow fiber bundle may be obtained through a washing process and then drying in a dryer, and the production method is not specified.

  A known method may be used for a method for producing a filter using a hollow fiber. For example, a hollow fiber membrane bundle is inserted into a cylindrical container having a fluid inlet / outlet port, a potting agent such as polyurethane is injected into both ends of the bundle to form a potting layer, and both ends are sealed. It can be produced by cutting and removing the potting agent, opening the end face of the hollow fiber, and attaching a header having a fluid inlet / outlet. By this method, the hollow fiber membrane bundle is filled in the container, a hollow fiber membrane inner chamber and a hollow fiber membrane outer chamber are formed, and has a fluid inlet / outlet leading to the hollow fiber membrane inner chamber and a fluid inlet / outlet leading to the hollow fiber membrane outer chamber. A hollow fiber membrane type concentration filter can be manufactured.

  Subsequently, the two-stage control of the present embodiment will be described.

  The two-stage control of the present embodiment includes the first step of passing the low concentration protein solution through the ascites concentration filter at the first flow rate, and when a predetermined amount or more is sent from the total amount of the low concentration protein solution. A second step of passing the low-concentration protein solution through the ascites filter at a second flow rate higher than the first flow rate.

  The switching from the first flow rate to the second flow rate is preferably performed when at least 1/4 of the total amount of the low-concentration protein solution is fed. Furthermore, the sieving coefficient of the protein that has passed through the ascites concentration filter is measured over time, and when an amount of 1/4 or more is fed and the sieving coefficient becomes 0.03 or less, the ascites inflow rate is set to the first rate. It is more preferable to switch from the flow rate to the second flow rate.

  The protein sieving coefficient can be calculated, for example, using a clinical refractometer (SUR-JE, Cat. No. 2733) manufactured by ATAGO. After dropping one or two drops on the prism surface of the refractometer, closing the daylighting plate, looking through the eyepiece, the position where the blue border line crosses the scale is the protein concentration. The sieve coefficient is calculated by dividing the measured protein concentration of the filtrate by the protein concentration of the raw ascites.

  The first flow rate is preferably 70 mL / min or less, more preferably 50 mL / min or less. In order to prevent protein leakage, low speed operation is preferable, but from the viewpoint of processing in a short time, 10 mL / min or less is not realistic, and 30 mL / min or more is realistic. On the other hand, the second flow rate is preferably 70 mL / min or less, more preferably 120 mL / min or less. The second flow rate is faster than the first flow rate, preferably 20 mL / min or more, more preferably 30 mL / min or more, and most preferably 40 mL / min or more faster than the first flow rate. In addition, when both the first and second flow rates are operated at low speed, there is an advantage that protein leakage can be prevented to the utmost limit, but it takes a long time to prepare a concentrated protein solution, and particularly a large amount When ascites is concentrated, it does not match the actual situation in the medical field. Specifically, a processing speed of about 3 L / h is required, and in order to realize this level of processing speed, the total value of the first flow velocity value and the second flow velocity value is 100 mL / min or more. It is preferable to become. Here, the sum of the first flow velocity value and the second flow velocity value is 100 mL / min or more, and the flow velocity difference between the first flow velocity and the second flow velocity is at least 20 mL / min or more. More preferred.

  As described above, by controlling the ascites inflow rate, the treatment can be completed in a short time while reducing protein loss due to the filtration concentration treatment. Specifically, when the above two-step control method is performed using bovine plasma under the same conditions, the protein concentration delivered from the filtration side outlet of the ascites concentration filter in the same processing time as compared with the conventional enforcement method Can be reduced to about half. Moreover, since the initial processing speed is low, it is possible to suppress the production of fever-causing substances, and the risk of fever after intravenous injection of ascites after concentration to the patient can be reduced.

  The protein concentration of the concentrated protein solution obtained by the method of this embodiment is 7 g / dL or more. If it is less than this concentration, even if it is administered to a patient, the effect of increasing the blood protein concentration of the patient is low, and the patient tends to accumulate ascites again. Moreover, it is preferable from the said effect that protein concentration shall be 10 g / dL or more.

  The method of this embodiment makes it possible to concentrate the protein concentration to about 5 times in a short time without an additional concentration step. The value obtained by dividing the concentrated protein concentration by the initial protein concentration is defined as the protein concentration ratio. The protein concentration ratio after the concentration reaches one fifth or less of the initial protein solution volume. When the value divided by the time (minutes) required for the above is defined as the protein concentration rate per hour, the protein concentration rate per hour is preferably 0.10 times / minute or more, preferably 0.15 times / minute. More preferably.

In the method of the present embodiment, the switching to the second step is preferably performed at the following timing. That is, the filter membrane area is A, the weight of the low-concentration protein solution processed in the first step is V1, the protein concentration of the low-concentration protein solution is C, the flow rate of the first step is Qb1, and the first step When the filtration flow rate is Qf1 and the ultrafiltration performance of the filter is F, it is preferable to carry out at a timing satisfying the following parameter (1).
103.7 ≦ −37log (A / V1) + log (Qb1 / V1) + 57log (F) −log (1 / C) −log (Qb1 / Qf1) ≦ 112.6 (1)
By using this parameter, it is possible to easily control the flow rate.

(Second Embodiment)
The ascites filtration and concentration apparatus 200 shown in FIG. 2 is configured to send ascites by the drop pressure of the liquid at each component. In order to perform liquid feeding using the drop pressure, in the ascites filtration and concentration apparatus 200, the drop pressure of the liquid at the inlet 3 a of the filter 3 for filtration is lower than the drop pressure of the liquid at the outlet 1 b of the storage container 1. The liquid drop pressure at the inlet 2 a of the recovery container 2 is lower than the liquid drop pressure at the concentrate outlet 4 b of the concentration filter 4. The liquid drop pressure at the ascites inlet 4 a of the concentration filter 4 is arranged to be lower than the liquid drop pressure at the filtrate outlet 3 c of the filter 3 for filtration.

  As a specific example of the above arrangement, as shown in FIG. 2, the filter 3 for filtration and the storage container 1 are arranged in a positional relationship such that the vertical position of the inlet 3a is lower than the vertical position of the outlet 1b. The The collection container 2 and the concentration filter 4 are arranged in a positional relationship such that the vertical position of the inlet 2a is lower than the vertical position of the concentrate outlet 4b. Further, the concentration filter 4 and the filtration filter 3 are arranged in such a positional relationship that the vertical position of the ascites inlet 4a is lower than the vertical position of the filtrate outlet 3c. With the arrangement as described above, liquid feeding using a drop pressure is performed in each of the flow paths 31 to 35, so that the pump 5 and the control device 41 (see FIG. 1) can be omitted. Note that the vertical position relationship of each part shown in FIG. 2 corresponds to the vertical position relation of each part of the actual ascites filtration concentrator 200 as it is. Further, in the ascites filtration and concentration apparatus 200, the same or equivalent components as those of the ascites filtration and concentration apparatus 100 described above are denoted by the same reference numerals and redundant description is omitted.

(Third embodiment)
In the ascites filtration concentrator 300 shown in FIG. 3, the liquid drop pressure at the ascites inlet 4 a of the concentration filter 4 is arranged to be equal to the liquid drop pressure at the filtrate outlet 3 c of the filter 3. Specifically, the concentration filter 4 and the filtration filter 3 are arranged in a positional relationship such that the vertical position of the ascites inlet 4a is the same height as the vertical position of the filtrate outlet 3c. With the arrangement as described above, it is possible to easily control the flow velocity only by adjusting the vertical position of the collection container 2. The positional relationship of other parts is the same as that of the ascites filtration and concentration apparatus 200 described above. Note that the vertical position relationship of each part shown in FIG. 3 corresponds to the vertical position relation of each part of the actual ascites filtration concentrator 300 as it is. In the ascites filtration and concentration apparatus 300, the same or equivalent components as those of the ascites filtration and concentration apparatus 200 described above are denoted by the same reference numerals, and redundant description is omitted.

  According to the ascites filtration concentration apparatus 100, 200, 300 and the method for producing a high-concentration protein solution described above, the concentration filter 4 has a specific range of ultrafiltration performance. Further, by controlling the flow rate through the concentration filter in at least two steps, the protein can be concentrated to a high magnification without loss. Further, since the treatment is completed in a short time, when the high-concentration protein solution obtained by the production method of the present embodiment is administered to the patient, the restraint time is shortened, which is preferable for the patient.

  In the method of the present embodiment, in the first step, the first step of sending the solution to the ascites concentration filter at the first flow rate, and the protein concentration in the solution at the inlet and the outlet of the ascites concentration filter are monitored. It is also preferable to perform control so that the second step is performed when the protein sieving coefficient drops to at least 0.03 or less.

  FIG. 4 is a diagram showing an ascites filtration and concentration device equipped with a refractometer as a device for monitoring the protein concentration in a solution. As shown in FIG. 4, a refractometer 50a is disposed in front of the inlet 4a of the concentrating filter 4, and a refractometer 50b is disposed on the outlet side of the concentrating filter 4, so that the solution at the inlet 4a and outlet 4c of the concentrating filter 4 is in solution. The protein concentration may be monitored and automatically controlled so that the second step is performed when the protein sieving coefficient drops to at least 0.03 or less.

  In the method of the present embodiment, in the first step, the first step of feeding the ascites concentration filter at the first flow rate, the weight of the low concentration protein solution is monitored, and the low concentration protein solution is monitored. It is also preferable to automatically control so that the second step is performed when a predetermined amount or more of the total amount is fed. Here, the predetermined amount is preferably at least 1/4 or more of the total amount of the low-concentration protein solution.

  FIG. 5 is a diagram showing an ascites filtration and concentration device equipped with a control device (for weight monitoring). As shown in FIG. 5, for example, a control device 60 for weight monitoring is installed in the storage container 1 to monitor the weight of the collected low-concentration protein solution that is the collected ascites in the storage container 1. What is necessary is just to control automatically so that a 2nd step may be performed when a predetermined amount or more is liquid-fed from the whole quantity. For example, the control device 60 may be a device that increases the height position of the storage container 1 and thereby increases the flow rate of the pump when the weight of the low concentration protein solution in the storage container 1 decreases by a certain amount. It is also possible to use a scale that is linked to a computer and that controls to increase the flow rate of the pump when the weight of the low concentration protein solution in the storage container 1 is reduced by a certain amount.

  EXAMPLES Hereinafter, although this invention is demonstrated further in detail according to an Example, this invention is not limited to these.

  The test method for ascites concentration performance is shown below. Bovine plasma adjusted to a total protein concentration of 3 g / dL (7 g / dL in Comparative Example 9) was used as simulated ascites (raw ascites), and 3 L was prepared. Since bovine plasma with no cellular components is used as simulated ascites, this simulated ascites can be regarded as having passed through a filter. And the filter 3 for filtration was implemented by the method which abbreviate | omitted. That is, the performance test of each concentration filter 54 according to the example and the comparative example was performed by the test apparatus 400 shown in FIG. The test apparatus 400 has a structure in which the first flow path 31 and the second flow path 32 are directly connected to the concentration filter 54 without using the filter 3 for filtration in the apparatus 100 shown in FIG. Polyvinyl chloride bags were used for the storage container 1 and the collection container 2, polyvinyl chloride tubes were used for the respective flow paths, roller pumps were used for the pump 5, and roller clamps were used as the control unit 15.

  Control of the inflow speed to the filter 54 for simulating ascites was performed by adjusting the pump 5. In Examples 1-12, ascites inflow rate is controlled to 30-70 mL / min until at least 1/4 volume of the low concentration protein solution stored in the storage container 1 is processed, After the treatment, the ascites inflow rate was adjusted to a flow rate of 70 to 120 mL / min, and simulated ascites was introduced into the concentration filter 54. Specifically, in the test apparatus 400 shown in FIG. 6, the pump 5 is operated at a rate of 30 − per minute until 0.75 L, which is a quarter amount of 3 L simulated ascites, is processed, that is, for about 10 to 30 minutes. After adjusting to 70 mL / min, simulated ascites was introduced into the concentration filter 54. The amount of liquid stored in the storage container 1 was measured by a gravimetric method, and if the liquid volume was 2.25 L or less, the pump 5 was adjusted to 50 to 120 mL / min per minute.

  Regarding the concentration target, for example, when the simulated ascites protein concentration is 3 g / dL, in order to obtain a protein concentration of about 15 g / dL as a concentrated protein solution, the protein concentration of the concentrated protein solution is 5 times the protein concentration of the liquid to be treated. The goal was to concentrate to a degree. In order to concentrate the protein concentration to about 5 times, the amount of the concentrated protein solution was adjusted to be 1/5 or less of the amount of the liquid to be treated. Specifically, the flow rate of the waste liquid discharged from the filtrate outlet 4 c of the concentration filter 54 and the amount of inflow into the collection container 2 installed above the concentration filter 54 are compressed by the flow path 35 of the roller clamp 15. While adjusting according to the degree, the simulated ascites was concentrated. At the stage where all 3 L of simulated ascites is introduced into the concentration filter 54, the roller pump is stopped and the amount of liquid collected in the collection container 2 is measured by a gravimetric method, and the amount of liquid is 1/5 or less of the liquid to be treated. If it has reached, the process is terminated here. On the other hand, if the amount of liquid did not reach 1/5 or less of the liquid to be treated, the circuit and the roller pump were rearranged, and the collection liquid in the collection container 2 was additionally concentrated. The additional concentrated amount was calculated based on the amount of remaining waste liquid required for the amount of the concentrated protein solution to reach 1/5 or less of the liquid to be treated. Each filter was evaluated based on whether additional concentration was necessary, the time required for the concentration process to reach the target (within 60 minutes), the protein concentration of the collected concentrated protein solution (15.0 g / dL or more) and the discharge. The protein concentration of the filtrate (waste liquid) (0.1 g / dL or less) was used as an index.

  Hereinafter, Examples 1 to 12 and Comparative Examples 1 to 11 will be described in more detail.

[Example 1]
18 parts by weight of a polysulfone resin (manufactured by Solvay, P-1700), 5 parts by weight of polyvinylpyrrolidone (hereinafter also referred to as PVP) (manufactured by Nippon Shokubai Co., Ltd., K-85N), N, N-dimethylacetamide (hereinafter also referred to as DMAC) A uniform film-forming stock solution consisting of 77 parts by weight was prepared. A 40% by weight aqueous solution of N, N-dimethylacetamide was extruded from the double nozzle simultaneously with the hollow inner solution, passed through a hood attached to block it from the outside, and coagulated with 50 ° C. water provided 30 cm below. It was immersed in a bath and wound up at a speed of 50 m / min. The resulting hollow fiber membrane was treated with a 20% by weight aqueous glycerin solution and then dried at 75 ° C. A hollow fiber membrane bundle was adjusted so as to have a membrane area of 1.5 m ² , loaded into a cylindrical container, and both ends were potted with a polyurethane resin to prepare a polysulfone hollow fiber membrane filter. The ultrafiltration performance of this filter was 107 mL / min / 200 mmHg. This filter was used as the above-described concentration filter 54, and ascites concentration operation was performed at an inflow rate of 30 mL / min until the time of ¼ volume processing of pseudo ascites, and thereafter, 70 mL / min. The concentration factor of 5 times or more could be reached, the concentration time was 57 minutes, and the protein concentration in the filtrate was 0.036 g / dL.

[Example 2]
A polysulfone concentration filter was produced in the same manner as in Example 1 except that the membrane area was 1.1 m ² . The ultrafiltration performance of this filter was 88 mL / min / 200 mmHg. When this filter was used as the above-described concentration filter, ascites concentration was performed at an inflow rate of 30 mL / min during the period up to ¼ volume processing of simulated ascites, and thereafter, 70 mL / min. The concentration rate could be doubled or more, the concentration time was 57 minutes, and the protein concentration in the filtrate was 0.035 g / dL.

[Example 3]
A polysulfone concentration filter was produced in the same manner as in Example 1 except that the membrane area was 2.1 m ² . The ultrafiltration performance of this filter was 134 mL / min / 200 mmHg. When this filter was used as the above-described concentration filter, ascites concentration was performed at an inflow rate of 30 mL / min during the period up to ¼ volume processing of simulated ascites, and thereafter, 70 mL / min. The concentration rate could be doubled or more, the concentration time was 57 minutes, and the protein concentration in the filtrate was 0.037 g / dL.

[Example 4]
A polysulfone concentration filter was produced in the same manner as in Example 1. The ultrafiltration performance of this filter was 107 mL / min / 200 mmHg. Using this filter as the above-mentioned concentration filter, the ascites concentration operation was performed at an inflow rate of 50 mL / min during the period up to ¼ volume processing of the simulated ascites, and thereafter, 100 mL / min. The concentration rate could be doubled or more, the concentration time was 37.5 minutes, and the protein concentration in the filtrate was 0.061 g / dL.

[Example 5]
In the same manner as in Example 2, a polysulfone concentration filter was produced. The ultrafiltration performance of this filter was 88 mL / min / 200 mmHg. Using this filter as the above-mentioned concentration filter, the ascites concentration operation was performed at an inflow rate of 50 mL / min during the period up to ¼ volume processing of the simulated ascites, and thereafter, 100 mL / min. The concentration rate could be doubled or more, the concentration time was 37.5 minutes, and the protein concentration in the filtrate was 0.060 g / dL.

[Example 6]
A filter for polysulfone concentration was produced in the same manner as in Example 3. The ultrafiltration performance of this filter was 134 mL / min / 200 mmHg. Using this filter as the above-mentioned concentration filter, the ascites concentration operation was performed at an inflow rate of 50 mL / min during the period up to ¼ volume processing of the simulated ascites, and thereafter, 100 mL / min. The concentration rate could be doubled or more, the concentration time was 37.5 minutes, and the protein concentration in the filtrate was 0.065 g / dL.

[Example 7]
A polysulfone concentration filter was produced in the same manner as in Example 1. The ultrafiltration performance of this filter was 107 mL / min / 200 mmHg. Using this filter as the above-mentioned concentration filter, the ascites concentration operation was performed at an inflow rate of 70 mL / min until the time of 1/4 volume treatment of simulated ascites, and thereafter, 120 mL / min. The concentration rate could be doubled or more, the concentration time was 29.5 minutes, and the protein concentration in the filtrate was 0.085 g / dL.

[Example 8]
In the same manner as in Example 2, a polysulfone concentration filter was produced. The ultrafiltration performance of this filter was 88 mL / min / 200 mmHg. Using this filter as the above-mentioned concentration filter, the ascites concentration operation was performed at an inflow rate of 70 mL / min until the time of 1/4 volume treatment of simulated ascites, and thereafter, 120 mL / min. The concentration rate could be doubled or more, the concentration time was 29.5 minutes, and the protein concentration in the filtrate was 0.082 g / dL.

[Example 9]
A filter for polysulfone concentration was produced in the same manner as in Example 3. The ultrafiltration performance of this filter was 134 mL / min / 200 mmHg. Using this filter as the above-mentioned concentration filter, the ascites concentration operation was performed at an inflow rate of 70 mL / min until the time of 1/4 volume treatment of simulated ascites, and thereafter, 120 mL / min. The concentration rate could be doubled or more, the concentration time was 29.5 minutes, and the protein concentration in the filtrate was 0.088 g / dL.

[Example 10]
A polysulfone concentration filter was produced in the same manner as in Example 1. The ultrafiltration performance of this filter was 150 mL / min / 200 mmHg. Using this filter as the above-mentioned concentration filter, the ascites concentration operation was performed at an inflow rate of 70 mL / min until the time of 1/4 volume treatment of simulated ascites, and thereafter, 120 mL / min. The concentration rate could be doubled or more, the concentration time was 29.5 minutes, and the protein concentration in the filtrate was 0.083 g / dL.

[Example 11]
A polysulfone concentration filter was produced in the same manner as in Example 1. The ultrafiltration performance of this filter was 85 mL / min / 200 mmHg. Using this filter as the above-mentioned concentration filter, the ascites concentration operation was performed at an inflow rate of 50 mL / min during the period up to ¼ volume processing of the simulated ascites, and thereafter, 100 mL / min. The concentration rate could be doubled or more, the concentration time was 37.5 minutes, and the protein concentration in the filtrate was 0.065 g / dL.

[Example 12]
A polysulfone concentration filter was produced in the same manner as in Example 1. The ultrafiltration performance of this filter was 110 mL / min / 200 mmHg. Using this filter as the above-mentioned concentration filter, the ascites concentration operation was performed at an inflow rate of 70 mL / min until the time of 1/4 volume treatment of simulated ascites, and thereafter, 120 mL / min. The concentration rate could be doubled or more, the concentration time was 29.5 minutes, and the protein concentration in the filtrate was 0.083 g / dL.

[Comparative Example 1]
A concentration filter was produced in the same manner as in Example 1. When this filter was used as the above-mentioned concentration filter and the ascites concentration operation was always performed at a constant inflow rate (100 mL / min), it was possible to reach a concentration ratio of 5 times or more without additional concentration, and the concentration time was 30 minutes. However, the protein concentration in the filtrate was 0.207 g / dL.

[Comparative Example 2]
A concentration filter was produced in the same manner as in Example 1. When this filter was used as the above-mentioned concentration filter and ascites was concentrated at a constant inflow rate (200 mL / min), it was possible to reach a concentration ratio of 5 times or more without additional concentration, and the concentration time was 15 minutes. However, the protein concentration in the filtrate was 0.498 g / dL.

[Comparative Example 3]
A concentration filter was produced in the same manner as in Example 1. When this filter was used as the above-described concentration filter, ascites concentration was performed at an inflow rate of 30 mL / min until the time of quasi-ascitic fluid 1/4 volume treatment, and thereafter, 50 mL / min. Although the concentration rate could be doubled or more, the concentration time was 70 minutes exceeding the limit time of 60 minutes, and the protein concentration in the filtrate was 0.030 g / dL.

[Comparative Example 4]
An ascites concentrator manufactured by Asahi Kasei Kuraray Medical Co., Ltd. having an ultrafiltration performance of 77 mL / min / 200 mmHg, AHF-UNH (polyacrylonitrile hollow fiber, 1.1 m ² ) was used. The ascites concentrator was diverted to the above-mentioned concentration filter, and an ascites concentration performance test was conducted. The ascitic fluid concentration operation was performed at an inflow rate of 50 mL / min until the time of treatment with a quasi-ascitic fluid of 1/4 volume, and thereafter 100 mL / min. When all the simulated ascites were processed and the amount of the collected liquid was measured, the target magnification was not reached, and the circuit and the roller pump were changed, and additional concentration of the collected liquid in the collection container was necessary.

[Comparative Example 5]
A uniform spinning dope comprising 18 parts by weight of PSf (manufactured by Solvay, P-1700), 4.5 parts by weight of PVP (manufactured by Nippon Shokubai Co., Ltd., K-85N) and 77.5 parts by weight of dimethylacetamide was prepared. Simultaneously with a hollow inner solution of DMAC 57% aqueous solution, it was extruded from a double nozzle and passed through a hood attached to block it from the outside, and immersed in a 75 ° C. coagulation bath made of water provided 90 cm below, and 40 m / min. Winded up at speed. The obtained hollow fiber membrane was treated with a 40% by weight glycerin aqueous solution and then dried at 80 ° C.
The hollow fiber membrane bundle was adjusted so that the membrane area was 2.0 m ² , loaded into a cylindrical container, and both ends were potted with a polyurethane resin to prepare a polysulfone hollow fiber membrane filter. The ultrafiltration performance of this filter was 170 mL / min / 200 mmHg.
Using this filter as the above-described concentration filter, ascites concentration operation was performed at an inflow rate of 50 mL / min until the time of quasi-ascitic fluid 1/4 treatment, and thereafter 100 mL / min. When all the simulated ascites were processed and the amount of the recovered solution was measured, the concentration time was 37.5 minutes. The protein concentration of the recovered solution was 3.0 g / dL, and the protein concentration of the target magnification was 15.0 g / d. dL was not reached. The protein concentration of the filtrate was also 2.951 g / dL.

[Comparative Example 6]
A concentration filter was produced in the same manner as in Example 1. The ultrafiltration performance of this filter was 107 mL / min / 200 mmHg. Using this filter as a filter for concentration as described above, ascites concentration was performed at an inflow rate of 30 mL / min until the sifting coefficient of simulated ascites reached 0.07, and thereafter, no additional concentration. The concentration rate of 5 times or more could be reached and the concentration time was 54 minutes, but the filtrate protein concentration was 0.210 g / dL.

[Comparative Example 7]
A polysulfone concentration filter was produced in the same manner as in Example 1. The ultrafiltration performance of this filter was 160 mL / min / 200 mmHg. Using this filter as the above-mentioned concentration filter, the ascites concentration operation was performed at an inflow rate of 70 mL / min until the time of 1/4 volume treatment of simulated ascites, and thereafter, 120 mL / min. The concentration rate could be doubled or more, and the concentration time was 29.5 minutes, but the protein concentration in the filtrate was 1.753 g / dL.

[Comparative Example 8]
A polysulfone concentration filter was produced in the same manner as in Example 1. The ultrafiltration performance of this filter was 80 mL / min / 200 mmHg. Using this filter as a filter for concentration as described above, the ascites concentration operation was performed at an inflow rate of 70 mL / min until the processing of 1/4 volume of simulated ascites, and thereafter 120 mL / min. After processing and measuring the amount of the recovered liquid, the target magnification was not reached, and the circuit and the roller pump were rearranged, and additional concentration of the recovered liquid in the recovery container was necessary.

[Comparative Example 9]
In the same manner as in Example 2, a polysulfone concentration filter was produced. The ultrafiltration performance of this filter was 110 mL / min / 200 mmHg. Using this filter as the aforementioned concentration filter, bovine plasma adjusted to a total protein concentration of 7 g / dL was used as simulated ascites, and 3 L was prepared. Since the sieving coefficient reached 0.03 when 1/10 volume of simulated ascites was processed, ascites concentration was performed at an inflow rate of 70 mL / min until then, and 120 mL / min. When the amount of the recovered liquid was measured, the target magnification was not reached, and the circuit and the roller pump were rearranged, and additional concentration of the recovered liquid in the recovery container was necessary.

[Comparative Example 10]
In the same manner as in Example 2, a polysulfone concentration filter was produced. The ultrafiltration performance of this filter was 110 mL / min / 200 mmHg. When this filter was used as the above-mentioned concentration filter, ascites concentration was performed at an inflow rate of 90 mL / min until the time of 1/4 volume treatment of simulated ascites, and thereafter, 120 mL / min. The concentration rate could be doubled or more, and the concentration time was 27.1 minutes, but the protein concentration in the filtrate was 0.108 g / dL.

[Comparative Example 11]
A concentration filter was produced in the same manner as in Example 2. The ultrafiltration performance of this filter was 110 mL / min / 200 mmHg. Using this filter as a filter for concentration as described above, ascites concentration was performed at an inflow rate of 70 mL / min until the sifting coefficient of simulated ascites reached 0.04, and thereafter, 120 mL / min. The concentration rate could be 5 times or more and the concentration time was 30.4 minutes, but the filtrate protein concentration was 0.113 g / dL.

Tables 1 and 2 show measurement conditions and measurement results of Examples 1 to 12 and Comparative Examples 1 to 11.

In Examples 1 to 12, the treatment time could be suppressed to 57 minutes while controlling the protein concentration delivered from the filtration side outlet of the ascites concentration filter to 100 mg / dL or less. On the other hand, in Comparative Examples 1 and 2 in which ascites inflow rates were 100 mL / min and 200 mL / min and controlled to a constant value over the entire processing time, and in Comparative Example 10 in which the inflow rate of the first step was set earlier, the filtrate side of the concentration filter The concentration of became 100 mg / dL or more.
In Comparative Example 3 in which the flow rate of the first step was controlled to 30 mL / min and the flow rate of the second step was controlled to 50 mL / min, the filtrate protein concentration was about the same as in Examples 1 to 3, but the processing time was 70 minutes. The time limit exceeded 60 minutes. Further, in Comparative Examples 4 and 8 using a filter having a low ultrafiltration performance, it was not possible to achieve concentration more than 5 times by one concentration, and additional concentration was required. Furthermore, in Comparative Examples 5 and 7 using filters with extremely high ultrafiltration performance, the protein was not sufficiently concentrated, and only a protein solution with a low protein concentration was obtained.
In Comparative Example 6 in which the timing of shifting to the second step is set at a time point when the sieve coefficient is 0.07, and in Comparative Example 11 in which the screen coefficient is set at 0.04, the protein concentration on the filtrate side is 100 mg / kg. dL or more. In Comparative Example 9 in which the protein concentration of ascites was set high, the concentration of 5 times or more could not be achieved by one concentration, and additional concentration was required.

  According to the present invention, in a method for concentrating a dilute protein solution such as ascites and obtaining a concentrated protein solution, the processing speed is not reduced due to clogging, and a high protein concentration is obtained without burden on the operator such as an additional concentration step. It becomes possible to provide a manufacturing method and a manufacturing apparatus of a high concentration protein solution that can obtain a concentrated protein solution.

DESCRIPTION OF SYMBOLS 1 ... Storage container, 1b ... Outlet (connection part of a storage container and a 1st flow path), 2 ... Recovery container, 2a ... Inlet (connection part of a recovery container and a 4th flow path) 3) Filtration filter, 3a ... Filtration filter inlet, 3b ... Filtration filter outlet, 3c ... Filtrate outlet (filter side outlet of ascites filter), 4,54 ... Concentration filter, 4a ... Ascites inlet (inlet of ascites concentration filter), 4b ... Concentrate outlet (exit of ascites concentrate filter), 4c ... Filtrate outlet (ascites concentration) Filter side outlet), 5 ... Pump (control means), 14, 15 ... Control section (control means), 31 ... First flow path, 32 ... Second flow path, 33. .. 3rd flow path, 34... 4th flow path, 35... 5th flow path, 41... Control device (control means), 50a, 5 b ... refractometer, (for weight monitoring) 60 ... control unit, 100, 200, 300 ... ascites filtration concentrator, 400 ... ascites concentrating testing apparatus.

Claims (16)

Aspirate the low-concentration protein solution through a circuit from a storage container in which the low-concentration protein solution is stored, using a polysulfone-based hollow fiber membrane type ascites with an ultrafiltration performance of 85 mL to 150 mL / min / 200 mmHg and a hydrophilic polymer. A first step of letting the solution pass through and letting out the filtrate from the filtration side outlet of the filter and sending out the high concentration protein solution from the outlet of the filter;
A second step of collecting the high concentration protein solution delivered from the outlet of the filter in a collection container;
Including
The first step includes
A first step of passing the low concentration protein solution through the ascites concentration filter at a first flow rate;
A second step of passing the low-concentration protein solution through the ascites concentration filter at a second flow rate faster than the first flow rate when a predetermined amount or more is sent from the total amount of the low-concentration protein solution;
A method for producing a high concentration protein solution, comprising: The membrane area of the filter is A, the weight of the low-concentration protein solution treated in the first step is V1, the protein concentration of the low-concentration protein solution is C, the flow rate of the first step is Qb1, and the first step 2. The method according to claim 1, wherein when the filtration flow rate in step 1 is Qf1 and the ultrafiltration performance of the filter is F, switching to the second step is performed at a timing satisfying the following (1).
103.7 ≦ −37log (A / V1) + log (Qb1 / V1) + 57log (F) −log (1 / C) −log (Qb1 / Qf1) ≦ 112.6 (1)   The method according to claim 1 or 2, wherein the switching from the first flow rate to the second flow rate is performed when at least 1/4 or more of the total amount of the low concentration protein solution is fed.   The method according to any one of claims 1 to 3, wherein the first flow rate is 70 mL / min or less and the second flow rate is 120 mL / min or less.   The method according to any one of claims 1 to 4, wherein the first flow rate is 50 mL / min or less and the second flow rate is 70 mL / min or less.   The sum of the first flow rate and the second flow rate is 100 mL / min or more, and the flow rate difference between the first flow rate and the second flow rate is at least 20 mL / min or more. The method of any one of these.   The method according to any one of claims 1 to 6, wherein in the first step, the protein concentration of the low concentration protein solution is 5 g / dL or less.   The method according to any one of claims 1 to 6, wherein in the first step, the protein concentration of the low concentration protein solution is 3 g / dL or less.   In the first step, the second step is started when a sieving coefficient of a protein in the high concentration protein solution delivered from the filtration side outlet of the ascites concentration filter is not more than a predetermined value. The method of any one of -8.   In the first step, the second step is started when a protein sieving coefficient in the high concentration protein solution delivered from the filtration side outlet of the ascites concentration filter is at least 0.03 or less. Item 10. The method according to any one of Items 1 to 9.   The method according to any one of claims 1 to 10, wherein in the first step, the protein concentration in the filtrate sent from the filtration-side outlet of the ascites concentration filter is 100 mg / dL or less.   The method according to any one of claims 1 to 11, wherein in the first step, the protein concentration of the high-concentration protein solution delivered from the outlet of the ascites concentration filter is 7 g / dL or more.   The method according to claim 1, wherein the circuit includes an ascites filter.   In the first step, the first step of feeding the ascites concentration filter to the ascites concentration filter at a first flow rate, the protein concentration in the solution at the inlet and the outlet of the ascites concentration filter, and monitoring the protein The method according to claim 1, wherein the second step is controlled when the coefficient has decreased to at least 0.03 or less.   In the first step, the first step of sending the liquid to the ascites concentration filter at a first flow rate, and monitoring the weight of the low concentration protein solution, a predetermined amount or more from the total amount of the low concentration protein solution is The method according to any one of claims 1 to 13, wherein the second step is controlled so as to be performed when the liquid is fed. Aspirate the low-concentration protein solution through a circuit from a storage container in which the low-concentration protein solution is stored, using a polysulfone-based hollow fiber membrane type ascites with an ultrafiltration performance of 85 mL to 150 mL / min / 200 mmHg and a hydrophilic polymer. A part having a first step of letting the solution pass through and letting out the filtrate from the filtration side outlet of the filter and sending out the high concentration protein solution from the outlet of the filter;
A portion having a second step of collecting the high-concentration protein solution delivered from the outlet of the filter in a collection container;
Including
The part having the first step comprises
A portion having a first step of passing the low concentration protein solution through the ascites concentration filter at a first flow rate;
A second step of passing the low-concentration protein solution through the ascites concentration filter at a second flow rate faster than the first flow rate when a predetermined amount or more is delivered from the total amount of the low-concentration protein solution. And
Including
The membrane area of the filter is A, the weight of the low-concentration protein solution treated in the first step is V1, the protein concentration of the low-concentration protein solution is C, the flow rate of the first step is Qb1, and the first step An apparatus for producing a high-concentration protein solution that switches to the second step at a timing that satisfies the following (1), where Qf1 is the filtration flow rate of step 1 and F is the ultrafiltration performance of the filter.
103.7 ≦ −37log (A / V1) + log (Qb1 / V1) + 57log (F) −log (1 / C) −log (Qb1 / Qf1) ≦ 112.6 (1)

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