TWI568461B - Method for producing high concentration protein solution and manufacturing device thereof - Google Patents

Description

Method and device for manufacturing high concentration protein solution

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

In the past, patients who are easily accumulating ascites such as cirrhosis or pleural effusion (hereinafter referred to as ascites) are used to increase the protein concentration in the blood of the patient by using protein in the ascites, and the following ascites filtration is concentrated and re-injected, that is, by using There are two kinds of filters such as a hollow fiber membrane, and the ascites discharged into the reservoir and discharged to the outside of the body is subjected to filtration concentration treatment to obtain a thick protein solution, which is injected into the patient (for example, refer to Patent Documents 1 and 2). ). The first filter of the two kinds of filters is a filter for removing cellular components such as cancer cells and blood cell components contained in the ascites, and a pore having a solute component such as moisture or protein which does not pass through the cellular component can be used. The film. On the other hand, another type of filter is a concentrated filter for concentrating protein from the ascites of a lean protein concentration, and a membrane which allows water, electrolyte, or the like to pass through without passing the protein component. In general, from the viewpoint of convenience, it is possible to adopt a method in which a cell component is filtered and separated by a filter, and a filter-separated ascites water is concentrated by a concentrator, and such a continuous device can be used.

On the other hand, as a method of suppressing protein leakage, in the field of hemodiafiltration, there is a method of controlling the filtration flow rate by using a device, thereby controlling the TMP of the membrane to achieve optimal diafiltration (for example, Patent Document 3).

[Previous Technical Literature] [Patent Literature]

[Patent Document 1] Japanese Patent Laid-Open Publication No. 2009-297242

[Patent Document 2] New Registration No. 2543466

[Patent Document 3] Japanese Patent Laid-Open Publication No. 2001-112863

The protein solution administered to the patient also has the purpose of increasing the protein concentration in the blood of the patient after administration, and therefore must be a certain high concentration, and it is necessary to concentrate to obtain a higher concentration ratio. When the protein concentration of the treated ascites is increased, the concentration filter is clogged by the protein, and the concentration efficiency is lowered, so that the concentration ratio of the target cannot be concentrated. In the above case, it is necessary to carry out a very complicated procedure in which the protein solution is temporarily recovered in a state where the concentration is insufficient, and the concentration is additionally carried out. As a method, it is necessary to replace the filter for concentration with a new filter depending on the situation, and therefore, it imposes a burden on the operator and is also economically disadvantageous. Further, in the case of the patient, since the treatment time is required for the addition, there is a disadvantage that the predetermined time until the administration of the protein solution becomes long, or when the concentration is added, the protein is wasted when the protein is recovered, and the administration amount is lowered.

On the other hand, patients with ascites storage can be roughly classified into: patients with hepatic ascites stored for diseases such as cirrhosis; and patients with cancerous ascites stored for cancers such as gastric cancer, ovarian cancer, and colon cancer. Previously, this treatment was mainly performed on patients with hepatic ascites. However, in recent years, the therapeutic effect of this treatment on patients with cancerous ascites has been continuously recognized, and the opportunities for patients with cancerous ascites have increased. However, cancerous ascites usually has a tendency to have a protein concentration richer than hepatic ascites, and therefore it is necessary to have an additional concentration step as described above.

Further, Patent Document 1 discloses a method of concentrating by a suction device provided on a waste liquid side of a concentrating filter, and providing a cathode pressure generating device downstream of the concentrating filter to suppress accumulation. Stacking of objects, but in this method, in order to open the settings The pressure release line in the middle of the suction device releases the suction pressure applied to the concentrator, which must be dealt with by the practitioner.

Further, when the ascites filtration and concentration treatment is performed, if the ascites inflow rate is low, the treatment time becomes long, and the patient's appointment time is prolonged and the burden is increased. On the other hand, if the inflow rate of the ascites is increased, there is a possibility that the protein useful on the filtrate side of the filter for ascites concentration leaks.

The object of the present invention is to provide a method and a device for producing a high-concentration protein solution, which is a method for producing a high-concentration protein solution by concentrating a thin protein solution such as ascites to obtain a thick protein solution. The method does not cause a decrease in the processing speed, and does not impose an additional concentration step on the burden on the implementer, and a thick protein solution having a higher protein concentration can be obtained.

The inventors of the present invention conducted intensive studies on the above problems, and as a result, found that the above-mentioned clogging is related to the ultrafiltration performance of the concentrating filter, and by using a concentrating filter having a specific ultrafiltration performance, the decrease in the concentration rate can be suppressed, and By controlling the liquid permeation rate to the concentration filter in at least two stages, the protein can be concentrated in a short time to a high magnification without loss, and the present invention has been completed.

Further, the liquid permeation rate is preferably controlled when the protein sieve coefficient or the original ascites weight is a fixed amount or less. However, it is difficult to measure the protein concentration or calculate the weight by frequently calculating the sieve coefficient during the application. On the other hand, it has been found through experiments that, as a factor for changing the numerical value of the sieve coefficient, not only the permeability of the filter itself, the membrane area or the amount of raw ascites treated, but also the value of the sieve coefficient is affected, thereby completing The invention of the parameters of the liquid permeation rate is simply controlled.

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

[1]

A method for producing a high concentration protein solution, comprising: In the first step, the low-concentration protein solution is passed through a line and the ultrafiltration performance is 85 mL to 150 mL/min/200 mmHg, and the hydrophilic polymer is given a polyfluorene-based hollow fiber membrane type. The ascites concentration filter sends the filtrate from the filter side outlet of the filter, and sends a high concentration protein solution from the outlet of the filter; in the second step, the high concentration protein solution sent from the outlet of the filter The first step includes: in the first step, the low-concentration protein solution is passed through the ascites concentration filter at a first flow rate; and the second step is from the total low-concentration protein solution. When the amount of the liquid is supplied at a specific amount or more, the low-concentration protein solution is passed through the ascites-concentrating filter at a second flow rate faster than the first flow rate.

[2]

[1] The method of [1], wherein the membrane area of the filter is A (m ² ), and the weight of the low-concentration protein solution treated in the first stage is V1 (kg), The protein concentration of the low-concentration protein solution is C (g/dL), the flow rate of the first stage is Qb1 (mL/min), and the filtration flow rate of the first stage is Qf1 (mL/min). When the ultrafiltration performance of the filter is F (mL/min/200 mmHg), the second stage is switched to when the following (1) is satisfied.

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 at a time point when at least 1/4 or more of the total amount of the low-concentration protein solution is supplied.

[4]

The method of any one of [1] to [3] wherein the first flow rate is 70 mL/min. Next, 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 method according to any one of [1] to [5] wherein a total value of the first flow rate and the second flow rate is 100 mL/min or more, and a difference in flow rate between the first flow rate and the second flow rate is at least 20 mL/min. the above.

[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]

The method according to any one of [1] to [8] wherein, in the first step, the sieve coefficient of the protein in the high-concentration protein solution sent from the filtration side outlet of the ascites-concentrating filter is a specific value In the following, the second stage described above is started.

[10]

The method according to any one of [1] to [9] wherein, in the first step, the protein of the protein in the high-concentration protein solution sent from the filtration side outlet of the ascites concentration filter is at least 0.03 In the following, the second stage described above is started.

[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 sent from the outlet of the ascites-concentrating filter is 7 g/dL or more.

[13]

The method of any one of [1] to [12] wherein the above line contains a filter for ascites filtration.

[14]

The method according to any one of [1] to [13] wherein, in the first step, the first step of controlling the ascites by the first flow rate to the ascites concentration filter is performed. The protein concentration in the solution at the inlet and outlet of the filter for concentration is carried out in the second stage at a time when the sieve coefficient of the protein is lowered to at least 0.03.

[15]

The method according to any one of [1] to [13] wherein, in the first step, the first step of controlling the lowering of the liquid to the ascites concentration filter by the first flow rate is performed as follows: The weight of the concentrated protein solution is subjected to the second stage at a time point when the total amount of the low-concentration protein solution is more than a predetermined amount.

[16]

A manufacturing apparatus for a high-concentration protein solution, comprising: a part comprising the first step, wherein the low-concentration protein solution passes through a line and has an ultrafiltration performance of 85 mL from a storage container in which a low-concentration protein solution is stored. 150mL/min/200mmHg and a hydrophilic polymer high-concentration hollow fiber membrane type ascites concentration filter, the filtrate is sent out from the filter side outlet of the filter, and a high-concentration protein solution is sent from the outlet of the filter; Included in the second step, the second step is to recover the high-concentration protein solution sent from the outlet of the filter into a recovery container, and the portion including the first step includes: a portion having a first stage, In the first stage, the low-concentration protein solution is passed through the ascites concentration filter at a first flow rate; and the second stage is a second stage, and the second stage is supplied from a total amount of the low-concentration protein solution to a specific amount or more. At a time point, the low-concentration protein solution is passed through the ascites concentration filter at a second flow rate faster than the first flow rate, and The membrane area of the filter set A (m ^2), via said first stage will be in a low concentration by weight of the above-described processing of the protein solution to V1 (kg), the protein concentration of the low concentration of the protein solution to C (g/dL), the flow rate of the first stage is set to Qb1 (mL/min), and the filtration flow rate of the first stage is set to Qf1 (mL/min), and the ultrafiltration performance of the filter is set to In the case of F (mL/min/200 mmHg), the second stage is switched to when the following (1) is satisfied.

103.7≦-37log(A/V1)+log(Qb1/V1)+57log(F)-log(1/C)-log(Qb1/Qf1)≦112.6(1)

According to the present invention, there can be provided a method and a manufacturing apparatus for producing a high-concentration protein solution which is a method for obtaining a concentrated protein solution by concentrating a thin protein solution such as ascites without causing clogging. The processing speed is lowered, and a thick protein solution having a high protein concentration can be obtained without burdening the implementer such as an additional concentration step.

1‧‧‧ storage container

1b‧‧‧Export (connection between storage container and first flow path)

2‧‧‧Recycling container

2a‧‧‧ entrance (connection between the recovery container and the fourth flow path)

3‧‧‧Filter filter

3a‧‧‧Inlet filter filter

3b‧‧‧Export of filter filter

3c‧‧‧ filtrate outlet (filter side outlet for ascites filtration)

4, 54‧‧‧ Concentration filter

4a‧‧‧ ascites inlet (inlet of ascites concentrate filter)

4b‧‧‧ Concentrate outlet (export of ascites concentrate filter)

4c‧‧‧ filtrate discharge port (filter side outlet for filter for ascites concentration)

5‧‧‧ pump (control agency)

14.15‧‧‧Control Department (Control Agency)

31‧‧‧1st flow path

32‧‧‧2nd flow path

33‧‧‧3rd flow path

34‧‧‧4th flow path

35‧‧‧5th flow path

41‧‧‧Control device (control mechanism)

50a, 50b‧‧ refractometer

60‧‧‧Control device (for weight monitoring)

100, 200, 300‧‧‧ ascites filtration and concentration device

400‧‧‧ Ascites concentration performance test device

Fig. 1 is a view showing a first embodiment of an ascites filtration and concentration device.

Fig. 2 is a view showing a second embodiment of the ascites filtration and concentration device.

Fig. 3 is a view showing a third embodiment of the ascites filtration and concentration device.

Fig. 4 is a view showing a ascites filtration concentrating device equipped with a refractometer.

Fig. 5 is a view showing an ascites filtration concentrating device including a control device (for weight monitoring).

Fig. 6 is a view showing a test apparatus for the concentration of ascites of a filter for concentration.

Hereinafter, the form for carrying out the present invention (hereinafter referred to as "this embodiment") will be described in detail. The present invention is not limited to the embodiments described below, and various modifications can be made without departing from the spirit and scope of the invention.

(First embodiment)

The configuration of the ascites filtration and concentration device 100 shown in FIG. 1 and an example of a method for obtaining a thick protein solution (high-concentration protein solution) by using the device 100 to filter and concentrate ascites (protein solution) is used as the first embodiment of the present invention. .

First, in the present embodiment, a low-concentration protein solution as a case of ascites taken in advance from a patient is stored in the storage container 1. Usually, the ascites is a low-concentration protein solution having a protein concentration of 5 g/dL or less. When the protein concentration exceeds 5 g/dL, the time required to form a dense layer of blood in the hollow fiber is short due to contamination, so that protein does not occur much even if it is initially passed at a relatively large flow rate. Leakage to the filtrate side, as a result, does not require the application of the method of the present invention. Similarly, in the case where the protein concentration exceeds 3 g/dL, although it is not more than 5 g/dL, a similar phenomenon occurs in a relatively short period of time, and thus there is a case where the method of the present invention is not necessarily applied. Therefore, the protein concentration of the low-concentration protein solution in the present embodiment is preferably 5 g/dL or less, more preferably 3 g/dL or less.

The outlet 1b of the storage container 1 is connected to the first flow path 31, and is connected to the inlet 3a of the filtration filter 3 via the first flow path 31. The ascites of the storage container 1 is supplied to the filtration filter 3 via the first flow path 31. In the filtration filter 3, since the cell component cannot pass through the filtration membrane, the filtered ascites containing the protein component from which the cell component has been removed is discharged from the filtrate outlet 3c of the filtration filter 3. The third flow path 33 is connected to the outlet 3b of the filter 3 for filtration, and a part of the ascites containing the cellular component is discharged to the outside of the apparatus 100 through the third flow path 33. The filtrate outlet 3c is connected to the ascites inlet of the concentration filter 4 via the second flow path 32. 4a, the filtered ascites discharged from the filtrate outlet 3c is introduced into the concentration filter 4 through the second flow path 32.

In the filter 4 for concentration, the moisture of the filtered ascites or the like is separated by filtration, and is discharged from the fifth flow path 35 connected to the filtrate discharge port 4c to the outside of the apparatus 100. Here, the liquid discharged from the filtrate discharge port 4c is preferably a low-concentration protein solution having a protein concentration of 100 mg/dL or less. The pore size of the filter 4 for concentration is smaller than the pore diameter of the filter 3 for filtration. Since the protein contained in the ascites after filtration is passed through the filter membrane and held in the filter for concentration 4, the protein concentration of the ascites is increased after the water is discharged, and the protein solution is concentrated as a concentrated protein solution. The concentrate outlet 4b of the filter 4 is discharged. The concentrated solution outlet 4b of the concentration filter 4 is connected to the inlet 2a of the recovery container 2 via the fourth flow path 34, and the concentrated protein solution discharged from the concentrated solution outlet 4b is recovered to the recovery container 2 via the fourth flow path 34. The concentrated protein solution recovered here has a protein concentration at least higher than that of the ascites of 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 any one as long as they can store the liquid. However, in general, a bag made of polyvinyl chloride can be used from the viewpoint of workability. The size of the container is determined according to the amount of ascites stored and the like.

The filter 3 for filtration is not particularly limited as long as it can separate a cell component from water, and a solute component such as an electrolyte or a protein. The structure, shape, and size of the filter are not limited as long as they have a ascites inlet and a filtered ascite outlet that can be connected to a flow path connected to the storage container 1 or the concentration filter 4. In addition, the material of the hollow fiber to be used for the filtration filter 3 is not particularly limited, and it is preferably a polyolefin or a polyether system such as polyethylene because the pore diameter is easily controlled during film formation and the chemical stability is excellent. , regenerated cellulose, polyvinyl alcohol, etc. Other materials may be contained in the hollow fiber materials exemplified above, or may be chemically modified. Generally, a hollow fiber membrane filter having a pore diameter of 0.2 μm or less and a protein permeability of 80% or more can be used.

In FIG. 1, the filtration filter 3 is used for the ascites filtration by the internal pressure filtration method. However, if the cell component can be separated by filtration, the external pressure filtration method may be used, and the third flow path 33 may be sealed. The filtering is performed by the end point method, and the third flow path 33 can be opened to be set as the sweeping mode.

The flow paths 31 to 35 constituting the apparatus of the first embodiment are not limited as long as they are connectable to the storage container 1, the recovery container 2, the filtration filter 3, and the concentration filter 4. Usually, a flexible tube made of polyvinyl chloride or the like can be used as the tube forming the flow paths 31 to 35.

Any mechanism can be used for the delivery of the ascites from the storage container 1 to the filtration filter 3. As an example, the pump 5 may be provided in the first flow path 31 as shown in FIG. 1 to perform liquid supply. A rotary pump or an infusion pump or the like can be generally used as the pump 5. Further, a liquid feeding pump may be added to the second flow path 32 and the third flow path 33.

In FIG. 1, the control unit 14 is provided in the middle of the fourth flow path 34, and the control unit 15 is provided 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 balance between the discharge amount of the filtrate of the concentration filter 4 and the amount of the concentrated protein solution can be adjusted by the control units 14 and 15, and the concentration ratio can be adjusted.

The control units 14 and 15 can control the balance between the amount of the filtrate discharged from the concentrated filter 4 and the amount of the liquid discharged from the concentrated liquid outlet 4b as long as it can control the ascites supplied to the concentration filter 4 and the amount of liquid discharged from the concentrated liquid outlet 4b. For anybody. When the control units 14 and 15 are exemplified, the flow path resistance may be adjusted by pressing the flow path such as a roll clamp to control the flow path, and the flow paths 34 and 35 may be controlled by applying a fixed negative pressure or positive pressure. The flow person can also be a device for controlling the flow rate such as a rotary pump or an infusion pump. The control units 14 and 15 may be only one of them as long as the balance between the amount of liquid discharged from the filter 4 for concentration and the amount of liquid discharged into the recovery container can be controlled.

The control device 41 controls the ascites in the first flow path 31 by controlling the driving of the pump 5. Flow rate (liquid volume per unit time). The control device 41 is, for example, a computer, and may also be an input terminal that allows the executor to input desired information about the control. Further, as the control units 14 and 15 are driven by the control signals from the control device 41, the flow rate of the flow paths 34 and 35 can be controlled by the control device 41.

The control device 41 controls the flow rate when the weight or protein concentration of the low-concentration protein solution becomes a value equal to or higher than a fixed value, for example, by monitoring the weight or protein concentration of the low-concentration protein solution.

In the apparatus 100 shown in FIG. 1, the first flow path 31 is connected below the filter filter 3, and the third flow path 33 is connected above the filter filter 3. However, even if it is reversely installed, The same effect can be obtained. Further, the second flow path 32 may be connected to an arbitrary position as long as it passes to the hollow fiber outer chamber portion of the filtration filter 3.

The ultrafiltration performance of the concentration filter 4 used in the present embodiment is 85 mL to 150 mL/min/200 mmHg. When the ultrafiltration performance is below this range, the amount of the filtrate discharged during concentration is lowered, and a sufficiently concentrated protein solution cannot be obtained. Further, as long as it is 95 mL/min/200 mmHg or more, the possibility of clogging is lower, which is preferable. If it is more than 150 mL/min/200 mmHg or more, the protein leaks out into the filtrate, and the protein concentration of a sufficient concentration cannot be obtained, which is not preferable.

The so-called ultrafiltration performance in the present embodiment is defined by the test shown below. It was prepared to adjust the protein concentration to 6 g/dL of bovine plasma, and the liquid was supplied to the concentration filter at a constant rate of 200 mL per minute by a rotary pump. At this time, the filtrate discharge port of the concentrator is in an open state. The line connected to the recovery liquid outlet of the concentration filter was pressed so as to adjust the pressure difference (hereinafter, also referred to as TMP (Transmembrane Pressure)) applied to the inside and outside of the filter to 200 mmHg. At this time, the volume per unit time of the filtrate discharged from the filtrate discharge port was measured. The TMP was calculated in the following manner.

TMP = (pressure on the inlet side of the filter + pressure on the outlet side of the filter) / 2 - filtrate side pressure

Further, in the present embodiment, a filter using a hollow fiber membrane is used from the viewpoint of concentration efficiency. The shape and size of the hollow fiber membrane are not particularly limited as long as they have the above-described ultrafiltration performance. The material is a polyfluorene system because it is easy to control the pore diameter at the time of film formation and is excellent in chemical stability. Since the polyfluorene-based polymer aromatic compound is particularly excellent in radiation resistance, it is also very strong in heat or chemical treatment, and is excellent in safety. Therefore, various film forming conditions can be selected and sterilized by radiation, which is particularly preferable as a film material for a ascites concentrator. In addition, the "~ system" has the following meanings: not only a homopolymer but also a copolymer with other monomers or a chemically modified substance.

Here, the polyfluorene-based polymer (hereinafter referred to as PSf) is a general term for a polymer compound having a hydrazone bond, and is not particularly limited. For example, the repeating unit is represented by the following formula (1) and formula (1). 2) A polyfluorene-based polymer represented by the formula (3), the formula (4), and the formula (5). A modified polymer having a substituent introduced into one of the aromatic rings may also be used. In terms of industrially readily available, it is preferred that the repeating unit is an aromatic polyfluorene-based polymer represented by the formula (1), the formula (2), and the formula (3), and particularly preferably having the formula (1) The cluster of chemical structures shown. The bisphenol type polyfluorene resin is commercially available under the trade name of "Udel (registered trademark)" from Solvay Advanced Polymers, and there are several types depending on the degree of polymerization, etc., but it is not particularly limited.

[Chemical 1]

The polyfluorene-based hollow fiber membrane in the present embodiment is hydrophilic by a hydrophilic polymer. The reason for this is that if only the polyfluorene-based polymer is used, the surface of the hollow fiber membrane becomes hydrophobic, and the protein is easily adsorbed on the surface, which is a cause of degrading the recovery performance of the protein. Examples of the hydrophilic polymer include polyvinylpyrrolidone (hereinafter referred to as PVP), polyethylene glycol, polyvinyl alcohol, and polypropylene glycol, but among them, the effect or safety of hydrophilization In terms of aspect, PVP is preferred. There are several types of PVP, and there are several types according to the molecular weight and the like. For example, K-15, 30, and 90 (all manufactured by ISP) of PVP are commercially available. The molecular weight (viscosity average molecular weight) of PVP used in the present 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%, based on the total amount of the polymer. When the content is 3% or less, the effect as a hydrophilizing agent is weakened, and when the content is more than 20%, the viscosity of the film forming solution is excessively increased, which is unsatisfactory in production.

A known dry-wet film forming technique can be applied to the method for producing a hydrophilized polyfluorene hollow fiber membrane. First, a polyfluorene-based polymer and a hydrophilic polymer such as polyvinylpyrrolidone are dissolved in a solvent common to both to prepare a uniform spinning dope. When the hydrophilic polymer is polyvinylpyrrolidone, the dimethylacetamide (hereinafter referred to as DMAC), dimethyl hydrazine, and N-methyl-2- can be mentioned as the above-mentioned common solvent. Pyrrolidone, dimethylformamide, cyclobutane, two A solvent such as an alkane or a solvent containing a mixture of two or more of the above solvents. Further, in order to control the pore diameter, an additive such as water may be added to the spinning dope.

In the production of the hollow fiber membrane, a tube in orifice type spinning head is used to discharge the spinning dope from the orifice of the spinning head into the air, and at the same time, the hollow inner liquid is discharged from the tube body into the air. The hollow inner liquid is used to solidify the spinning dope, and water or a coagulating liquid mainly composed of water can be used. The hollow inner liquid is not particularly limited as long as it depends on the properties such as the ultrafiltration performance of the hollow fiber membrane to be targeted, and generally cannot be used. Generally, a mixed solution of a solvent and water used in the spinning dope can be preferably used. For example, a 0 to 65 wt% DMAC aqueous solution or the like can be used as the hollow internal liquid. The spinning dope discharged from the spinning head together with the hollow inner liquid migrates in the vacant portion, and is introduced into a coagulation bath mainly composed of water provided in the lower portion of the spinning head, and is immersed to complete solidification. Thereafter, the solidified hollow fiber is subjected to a washing step or the like, and the hollow fiber membrane in a wet state is taken up by a coiler to obtain a bundle of hollow fiber membranes, followed by drying. Alternatively, the hollow fiber bundle may be obtained by a washing step and then drying in a dryer, and the manufacturing method is not specified.

The method for producing the filter using the hollow fiber may be any known method. For example, it can be manufactured by inserting a hollow fiber membrane bundle into a cylindrical container having a fluid inlet and outlet, injecting a potting agent such as polyurethane into a potting layer at both ends, and sealing the both ends, and then cutting The excess potting agent after hardening is removed to open the end face of the hollow fiber, and a header having a fluid inlet and outlet is installed. By this method, the hollow fiber membrane bundle can be manufactured to be filled in a container to form a hollow fiber membrane inner chamber and a hollow fiber membrane outer chamber. And a hollow fiber membrane type concentration filter having a fluid inlet and outlet to the inner chamber of the hollow fiber membrane and a fluid inlet and outlet to the outer chamber of the hollow fiber membrane.

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

The two-stage control of the present embodiment includes a first stage in which the low-concentration protein solution is passed through the ascites-concentrating filter at a first flow rate, and in the second stage, the liquid is supplied in a specific amount or more from the total amount of the low-concentration protein solution. At the time point, the low concentration protein solution was passed through the ascites concentration filter at a second flow rate faster than the first flow rate.

The switching from the first flow rate to the second flow rate is preferably performed at a time point of at least 1/4 or more of the total amount of the liquid-feeding low-concentration protein solution. Further, the sieve coefficient of the protein passing through the filter for ascites concentration is measured over time, and it is more preferable to switch the ascites inflow rate from the first flow rate to the second time point when the liquid supply amount is 1/4 or more and the sieve coefficient is 0.03 or less. Flow rate.

The protein sieve coefficient can be calculated, for example, using a clinical refractometer (SUR-JE, Cat. No. 2733) manufactured by ATAGO Corporation. Drop the sample 1 or 2 drops on the surface of the refractometer, close the lighting plate, and observe the eyepiece. The boundary line of the blue crosses the scale to become the protein concentration. The sieve coefficient was calculated by dividing the protein concentration of the filtrate obtained by the measurement by the protein concentration of the ascites.

The first flow rate is preferably 70 mL/min or less, more preferably 50 mL/min or less. In order to prevent leakage of protein, it is preferred to operate at a low speed. However, from the viewpoint of processing in a short period of time, 10 mL/min or less is not realistic, and the actual value is 30 mL/min or more. 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 faster than the first flow rate, more preferably 30 mL/min or more, and most preferably 40 mL/min or more. Further, when both the first and second flow rates are set to be low-speed operation, there is an advantage that leakage of protein can be prevented from reaching the limit. On the other hand, it takes a long time to produce a concentrated protein solution, particularly in the case of concentrating a large amount of ascites. At the same time, it is inconsistent with the actual situation at the medical site. Specifically, a processing speed of about 3 L/h is required, and in order to achieve the processing speed of the level, the first flow rate value and the second flow rate value are preferable. The total value is 100 mL/min or more. Here, it is more preferable that the total value of the first flow rate value and the second flow rate value is 100 mL/min or more, and the difference in flow rate between the first flow rate and the second flow rate is at least 20 mL/min or more.

As described above, the loss of protein due to the filtration concentration treatment can be reduced by controlling the inflow speed of the ascites, and the treatment can be completed in a short time. Specifically, when the above-described two-stage control method is carried out using the same conditions of bovine plasma, the protein concentration sent from the filtration side outlet of the ascites-concentrating filter can be suppressed to about half by the same treatment time as the previous method. Further, since the initial processing speed is low, it is possible to suppress the generation of the heat generating substance, and it is possible to reduce the risk of heat generation after the ascites filtered and concentrated is intravenously injected into the patient.

The protein concentration of the concentrated protein solution obtained by the method of the present embodiment is 7 g/dL or more. If the concentration is not reached, there is a drawback in that even if administered to a patient, the effect of increasing the protein concentration in the blood of the patient is low, and the patient is likely to store ascites or the like again. Further, in view of the above effects, the protein concentration is preferably 10 g/dL or more.

In the method of the present embodiment, there is no additional concentration step, and the protein concentration can be concentrated up to about 5 times in a short time. The value obtained by dividing the concentrated protein concentration by the initial protein concentration is defined as the protein concentration ratio, and the protein concentration ratio is divided by the liquid amount of the concentrated protein solution to reach one-fifth of the initial protein solution. The value obtained in the following time (minutes) is set to the protein concentration ratio per unit time, and the protein concentration ratio per unit time is preferably 0.10 times/min or more, more preferably 0.15 times/min or more.

In the method of the present embodiment, the switching to the second stage is preferably performed at the following timing. That is, it is preferable to set the membrane area of the filter to A (m ² ), and to set the weight of the low-concentration protein solution to be treated in the first stage to V1 (kg), and to set the protein concentration of the low-concentration protein solution. Set C (g/dL), set the flow rate of the first stage to Qb1 (mL/min), set the filtration flow rate of the first stage to Qf1 (mL/min), and set the ultrafiltration performance of the filter to F. In the case of (mL/min/200 mmHg), the second stage is performed at the time when the parameters of the following (1) are satisfied.

103.7≦-37log(A/V1)+log(Qb1/V1)+57log(F)-log(1/C)-log(Qb1/Qf1)≦112.6(1)

By using this parameter, the control of the liquid permeation speed can be simplified.

(Second embodiment)

The ascites filter concentrating device 200 shown in Fig. 2 is a person who transports ascites using the difference pressure of the liquid in each of the constituent parts. In order to carry out the liquid supply using the drop pressure, in the ascites filter concentrating device 200, the drop pressure of the liquid in the inlet 3a of the filter 3 for filtration becomes lower than the drop pressure of the liquid in the outlet 1b of the storage container 1. The arrangement is performed such that the drop pressure of the liquid in the inlet 2a of the recovery container 2 becomes lower than the drop pressure of the liquid in the concentrate outlet 4b of the concentration filter 4. Moreover, the drop pressure of the liquid in the ascites inlet 4a of the filter 4 for concentration is set to be lower than the drop pressure of the liquid in the filtrate outlet 3c of the filter 3 for filtration.

As a specific example of the above arrangement, as shown in FIG. 2, the filter filter 3 and the storage container 1 are disposed in a positional relationship such that the upper and lower positions of the inlet 3a are lower than the upper and lower positions of the outlet 1b. The recovery container 2 and the concentration filter 4 are disposed in a positional relationship such that the upper and lower positions of the inlet 2a are lower than the upper and lower positions of the concentrate outlet 4b. Moreover, the filtration filter 4 and the filtration filter 3 are arranged in a positional relationship such that the upper and lower positions of the ascites inlet 4a are lower than the upper and lower positions of the filtrate outlet 3c. By the above arrangement, the liquid supply by the drop pressure is performed in each of the flow paths 31 to 35, and therefore the pump 5 or the control device 41 (see Fig. 1) may be omitted. Furthermore, the positional relationship between the upper and lower parts of each part shown in FIG. 2 directly corresponds to the upper and lower positional relationship of each part of the actual ascites filtration and concentration apparatus 200. In the ascites filter concentrating device 200, the same or equivalent components as those of the ascites filter concentrating device 100 are denoted by the same reference numerals, and the description thereof will not be repeated.

(Third embodiment)

In the ascites filtration and concentration device 300 shown in FIG. 3, the ascites of the filter 4 for concentration is used. The difference pressure of the liquid in the inflow port 4a is set to be equal to the difference in the pressure of the liquid in the filtrate outlet 3c of the filter filter 3. Specifically, the filtration filter 4 and the filtration filter 3 are disposed in a positional relationship such that the upper and lower positions of the ascites inlet 4a are at the same height as the upper and lower positions of the filtrate outlet 3c. With the above configuration, the following effect can be obtained: the flow rate can be simply controlled by adjusting only the upper and lower positions of the recovery container 2. The positional relationship of the parts other than the above is the same as that of the above-described ascites filter concentrating device 200. Furthermore, the positional relationship between the upper and lower parts of each part shown in FIG. 3 directly corresponds to the upper and lower positional relationship of each part of the actual ascites filtration and concentration device 300. In the ascites filter concentrating device 300, the same or equivalent components as those of the ascites filter concentrating device 200 are denoted by the same reference numerals, and the description thereof will not be repeated.

According to the method for producing the ascites filtration and concentration devices 100, 200, and 300 and the high-concentration protein solution described above, the concentration filter 4 has a specific range of ultrafiltration performance, so that the concentration rate can be suppressed from decreasing, and at least 2 The stage controls the liquid permeation rate to the concentration filter, so that the protein can be concentrated to a high rate without loss. In addition, when the treatment is completed in a short period of time, when a high-concentration protein solution obtained by the production method of the present embodiment is administered to a patient, the restraining time is shortened, which is also preferable for the patient.

In the method of the present embodiment, it is preferable to perform the control in the first step of performing the first stage of supplying the liquid to the ascites concentration filter at the first flow rate, and monitoring the inlet of the ascites concentration filter. The protein concentration in the solution with the outlet is subjected to the second stage at a time point when the protein sieve coefficient falls below at least 0.03.

Fig. 4 is a view showing an ascites filtration concentrating device having a refractometer as a device for monitoring the protein concentration in a solution. As shown in Fig. 4, the refractometer 50a is disposed on the near side of the inlet 4a of the concentration filter 4, and the refractometer 50b is disposed on the outlet side of the concentration filter 4, and the control can be automatically performed as follows: The protein concentration in the solution of the inlet 4a and the outlet 4c of the concentration filter 4 is reduced to at least 0.03 in the protein sieve coefficient. The second phase is performed at the following time points.

Further, in the method of the present embodiment, it is preferable that the first step is automatically controlled in such a manner that the first stage of supplying the liquid to the ascites concentration filter at the first flow rate is performed, and the low concentration protein is monitored. The weight of the solution is subjected to the second stage at a time point when the total amount of the low-concentration protein solution is more than a certain amount. Here, the specific amount is preferably at least 1/4 or more of the total amount of the low-concentration protein solution.

Fig. 5 is a view showing an ascites filtration concentrating device including a control device (for weight monitoring). As shown in FIG. 5, for example, a control device 60 for weight monitoring is provided in the storage container 1, and the control can be automatically performed as follows: monitoring the low-concentration protein solution as the ascites taken in the storage container 1. The weight is subjected to the second stage at a time point when the total amount of the low-concentration protein solution is more than a predetermined amount. The control device 60 can be a device that is controlled in such a manner that, for example, if the weight of the low-concentration protein solution in the storage container 1 is reduced by a certain amount, the height position of the storage container 1 is increased to increase the flow rate of the pump, and It is a device which is a scale that is linked to a computer and is controlled in such a manner as 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, the present invention will be described in more detail based on examples, but the present invention is not limited thereto.

Hereinafter, a method of testing the concentration performance of ascites is shown. 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 the cell-free bovine plasma is used as the simulated ascites, the simulated ascites can be regarded as passing through the filter. Therefore, it is judged that the filter is omitted by omitting the filter even if the filter is omitted in the test. Implementation. That is, the performance test of each of the concentration filters 54 of the comparative example and the comparative example was carried out by the test apparatus 400 shown in FIG. The test apparatus 400 has a configuration in which the first flow path is made in the apparatus 100 shown in FIG. 31 and the second flow path 32 are directly connected to the concentration filter 54 without passing through the filtration filter 3. In the storage container 1 and the recovery container 2, a bag made of polyvinyl chloride is used, and a pipe made of polyvinyl chloride is used for each flow path, a pump 5 is used as a rotary pump, and a roller clamp is used as the control unit 15.

The control of the inflow velocity of the ascites to the concentration filter 54 is performed by adjusting the pump 5. In Examples 1 to 12, the ascites inflow rate was controlled to 30 to 70 mL/min until the treatment of the low-concentration protein solution stored in the storage container 1 was at least 1/4, and the treatment amount was adjusted to adjust the ascites. The inflow velocity was introduced into the concentration filter 54 at a flow rate of 70 to 120 mL/min. Specifically, in the test apparatus 400 shown in FIG. 6, the period of 0.75 L which is 1/4 of the simulated ascites of 3 L is treated, that is, about 10 to 30 minutes, and the ascites inflow rate becomes 30-70 mL per minute. The pump 5 is adjusted in the manner of /min, and the simulated ascites is introduced into the concentration filter 54. The amount of liquid stored in the storage container 1 is measured by the gravimetric method. When the amount of liquid is 2.25 L or less, the pump 5 is adjusted so that the aspirated inflow rate becomes 50-120 mL/min per minute.

In the case of the concentration target, for example, when the simulated ascites protein concentration is 3 g/dL, it is set as follows: In order to obtain a protein concentration of about 15 g/dL as a thick protein solution, concentration is performed until the protein concentration of the thick protein solution becomes The protein concentration of the treatment solution is about 5 times. In order to concentrate the protein concentration to about 5 times, the liquid amount of the thick protein solution is adjusted to be one-fifth or less of the liquid amount of the liquid to be treated. Specifically, the flow rate of the waste liquid discharged from the filtrate discharge port 4c of the concentration filter 54 and the recovery container 2 provided above the concentration filter 54 are adjusted according to the degree of compression of the flow path 35 by the roll holder 15. The inflow amount was simulated while the ascites was concentrated. When all of the simulated ascites 3L is introduced into the concentration filter 54, the rotary pump is stopped, and the amount of liquid recovered by the recovery container 2 is measured by a gravimetric method. If the amount of liquid reaches one-fifth or less of the liquid to be treated, The processing is completed at this time. On the other hand, if the amount of liquid does not reach one-fifth or less of the liquid to be treated, the line and the rotary pump are rearranged, and the concentrated liquid in the recovery container 2 is additionally concentrated. The additional concentration is calculated to calculate the amount of liquid in the thick protein solution. The amount of remaining waste liquid necessary for one-fifth or less of the chemical liquid is additionally concentrated based on this. The evaluation index of each filter is as follows: whether it is necessary to add additional concentration; the time taken for the target concentration treatment (within 60 minutes), the protein concentration of the concentrated thick protein solution (15.0 g/dL or more), and the discharged filtrate Protein concentration of (waste liquid) (0.1 g/dL or less).

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

[Example 1]

18 parts by weight of polyfluorene resin (P-1700, manufactured by Solvay Co., Ltd.), polyvinylpyrrolidone (hereinafter also referred to as PVP) (manufactured by Nippon Shokubai Co., Ltd., K-85N), 5 parts by weight, N, N-di 77 parts by weight of a uniform film-forming stock solution of methyl acetamide (hereinafter also referred to as DMAC). 40% by weight of N,N-dimethylacetamide aqueous solution and hollow inner liquid were simultaneously extruded from the double spinning nozzle, passed through a cover installed to isolate the outside, and immersed in a 50 ° C set below 30 cm. In the coagulation bath of water, it was taken up at a speed of 50 m/min. The obtained hollow fiber membrane was treated with a 20% by weight aqueous glycerin solution, and then dried at 75 °C. The hollow fiber membrane bundle was adjusted so that the membrane area became 1.5 m ² , and it was packed in a cylindrical container, and the both ends were potted by the polyurethane resin to produce a polyfluorene hollow fiber membrane filter. The ultrafiltration performance of the filter was 107 mL/min/200 mmHg. This filter was used as the concentration filter 54 as described above, and the ascites concentration operation was performed at an inflow rate of 70 mL per minute during the period of 1/4 of the simulated ascites, and the ascites concentration operation was performed at an inflow rate of 70 mL per minute. Concentrated to a concentration ratio of 5 times or more, the concentration time was 57 minutes, and the protein concentration in the filtrate became 0.036 g/dL.

[Embodiment 2]

A polyfluorene-concentrating 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 the filter was 88 mL/min/200 mmHg. This filter was used as the above-mentioned filter for concentration, and the ascites concentration operation was performed at a flow rate of 70 mL per minute during the period of 1/4 of the simulated ascites, and the concentration of the ascites was further performed at an inflow rate of 70 mL per minute. When the concentration ratio was 5 times or more, the concentration time was 57 minutes, and the protein concentration in the filtrate was 0.035 g/dL.

[Example 3]

A polyfluorene-concentrating 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 the filter was 134 mL/min/200 mmHg. This filter was used as the above-mentioned filter for concentration, and the ascites concentration operation was performed at a flow rate of 70 mL per minute during the period of 1/4 of the simulated ascites, and the concentration of the ascites was further performed at an inflow rate of 70 mL per minute. The concentration ratio was 5 times or more, the concentration time was 57 minutes, and the protein concentration in the filtrate was 0.037 g/dL.

[Example 4]

A polypene concentration filter was produced in the same manner as in Example 1. The ultrafiltration performance of the filter was 107 mL/min/200 mmHg. This filter is used as the above-mentioned filter for concentration, and the ascites concentration operation is performed at a flow rate of 100 mL per minute during the period of 1/4 of the simulated ascites, and then the ascites concentration operation is performed at an inflow rate of 100 mL per minute. On the other hand, the concentration ratio of 5 times or more was obtained, the concentration time was 37.5 minutes, and the protein concentration in the filtrate was 0.061 g/dL.

[Example 5]

A polypene concentration filter was produced in the same manner as in Example 2. The ultrafiltration performance of the filter was 88 mL/min/200 mmHg. This filter is used as the above-mentioned filter for concentration, and the ascites concentration operation is performed at a flow rate of 100 mL per minute during the period of 1/4 of the simulated ascites, and then the ascites concentration operation is performed at an inflow rate of 100 mL per minute. The concentration ratio was 5 times or more, the concentration time was 37.5 minutes, and the protein concentration in the filtrate was 0.060 g/dL.

[Embodiment 6]

A polypene concentration filter was produced in the same manner as in Example 3. The ultrafiltration performance of the filter was 134 mL/min/200 mmHg. This filter is used as the above-mentioned filter for concentration, and it is 50 mL per minute during the period of processing 1/4 of the simulated ascites, and thereafter The ascites concentration operation was performed at an inflow rate of 100 mL per minute. As a result, it was possible to achieve a concentration ratio of 5 times or more without additional concentration, the concentration time was 37.5 minutes, and the protein concentration in the filtrate was 0.065 g/dL.

[Embodiment 7]

A polypene concentration filter was produced in the same manner as in Example 1. The ultrafiltration performance of the filter was 107 mL/min/200 mmHg. This filter was used as the above-mentioned filter for concentration, and the ascites concentration operation was performed at a flow rate of 120 mL per minute during the period of 1/4 of the simulated ascites, and then the ascites concentration operation was performed at an inflow rate of 120 mL per minute. The concentration ratio was 5 times or more, the concentration time was 29.5 minutes, and the protein concentration in the filtrate was 0.085 g/dL.

[Embodiment 8]

A polypene concentration filter was produced in the same manner as in Example 2. The ultrafiltration performance of the filter was 88 mL/min/200 mmHg. This filter was used as the above-mentioned filter for concentration, and the ascites concentration operation was performed at a flow rate of 120 mL per minute during the period of 1/4 of the simulated ascites, and then the ascites concentration operation was performed at an inflow rate of 120 mL per minute. The concentration ratio was 5 times or more, the concentration time was 29.5 minutes, and the protein concentration in the filtrate was 0.082 g/dL.

[Embodiment 9]

A polypene concentration filter was produced in the same manner as in Example 3. The ultrafiltration performance of the filter was 134 mL/min/200 mmHg. This filter was used as the above-mentioned filter for concentration, and the ascites concentration operation was performed at a flow rate of 120 mL per minute during the period of 1/4 of the simulated ascites, and then the ascites concentration operation was performed at an inflow rate of 120 mL per minute. The concentration ratio was 5 times or more, the concentration time was 29.5 minutes, and the protein concentration in the filtrate was 0.088 g/dL.

[Embodiment 10]

A polypene concentration filter was produced in the same manner as in Example 1. This filter The ultrafiltration performance is 150 mL/min/200 mmHg. This filter was used as the above-mentioned filter for concentration, and the ascites concentration operation was performed at a flow rate of 120 mL per minute during the period of 1/4 of the simulated ascites, and then the ascites concentration operation was performed at an inflow rate of 120 mL per minute. The concentration ratio was 5 times or more, the concentration time was 29.5 minutes, and the protein concentration in the filtrate was 0.083 g/dL.

[Example 11]

A polypene concentration filter was produced in the same manner as in Example 1. The ultrafiltration performance of this filter is 85 mL/min/200 mmHg. This filter is used as the above-mentioned filter for concentration, and the ascites concentration operation is performed at a flow rate of 100 mL per minute during the period of 1/4 of the simulated ascites, and then the ascites concentration operation is performed at an inflow rate of 100 mL per minute. The concentration ratio was 5 times or more, the concentration time was 37.5 minutes, and the protein concentration in the filtrate was 0.065 g/dL.

[Embodiment 12]

A polypene concentration filter was produced in the same manner as in Example 1. The ultrafiltration performance of the filter is 110 mL/min/200 mmHg. This filter was used as the above-mentioned filter for concentration, and the ascites concentration operation was performed at a flow rate of 120 mL per minute during the period of 1/4 of the simulated ascites, and then the ascites concentration operation was performed at an inflow rate of 120 mL per minute. The concentration ratio was 5 times 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 filter for concentration was produced in the same manner as in Example 1. This filter is used as the above-mentioned filter for concentration, and the ascites concentration operation is always performed at a fixed inflow rate (100 mL per minute). As a result, it is possible to achieve a concentration ratio of 5 times or more without additional concentration, and the concentration time is 30 minutes, but the filtrate The protein concentration in the medium became 0.207 g/dL.

[Comparative Example 2]

A filter for concentration was produced in the same manner as in Example 1. Use this filter as In the above-mentioned filter for concentration, the ascites concentration operation is always performed at a fixed inflow rate (200 mL per minute), and as a result, it is possible to achieve a concentration ratio of 5 times or more without additional concentration, and the concentration time is 15 minutes, but the protein concentration in the filtrate becomes 0.498 g/dL.

[Comparative Example 3]

A filter for concentration was produced in the same manner as in Example 1. The filter was used as the concentration filter, and the ascites concentration operation was performed at a flow rate of 50 mL per minute during the period of 1/4 of the simulated ascites, and then the ascites concentration operation was performed at an inflow rate of 50 mL per minute. On the other hand, the concentration ratio of 5 times or more was obtained, but the concentration time was 70 minutes exceeding the time limit of 60 minutes, and the protein concentration in the filtrate was 0.030 g/dL.

[Comparative Example 4]

Ascites concentrator manufactured by Asahi Kasei Kuraray Medical Co., Ltd., which has an ultrafiltration performance of 77 mL/min/200 mmHg, is AHF-UNH (polyacrylonitrile hollow fiber, 1.1 m ² ). The ascites concentrator was used as the above-mentioned concentration filter to carry out an ascites concentration performance test. During the period of treatment of 1/4 of the simulated ascites, the ascites concentration operation was performed at an inflow rate of 100 mL per minute at 50 mL per minute. All the simulated ascites were treated, and the amount of the recovered liquid was measured. As a result, the target magnification was not reached, and it was necessary to rearrange the line and the rotary pump to perform additional concentration of the recovered liquid in the recovery container.

[Comparative Example 5]

A spinning dope containing 18 parts by weight of PSf (manufactured by Solvay Co., Ltd., P-1700), 4.5 parts by weight of PVP (K-85N, manufactured by Nippon Shokubai Co., Ltd.), and 77.5 parts by weight of dimethylacetamide was prepared. The DMAC 57% aqueous solution and the hollow internal liquid were simultaneously extruded from the double spinning nozzle, passed through a cover installed to isolate the outside, and immersed in a coagulation bath containing 75 ° C of water disposed below 90 cm, at 40 m/min. The speed is taken up. The obtained hollow fiber membrane was treated with a 40% by weight aqueous glycerin solution, and then dried at 80 °C.

The hollow fiber membrane bundle was adjusted so that the membrane area was 2.0 m ² , and it was packed in a cylindrical container, and the polystyrene resin was potted at both ends to produce a polyfluorene hollow fiber membrane filter. The ultrafiltration performance of the filter is 170 mL/min/200 mmHg.

This filter was used as the above-mentioned filter for concentration, and the ascites concentration operation was performed at an inflow rate of 100 mL per minute during the period of time to 1/4 of the simulated ascites. All the simulated ascites were treated, and the amount of the recovered liquid was measured. As a result, the concentration time was 37.5 minutes, but the protein concentration of the recovered liquid was 3.0 g/dL, and the protein concentration of the target magnification was 15.0 g/dL. Further, the protein concentration of the filtrate also became 2.951 g/dL.

[Comparative Example 6]

A filter for concentration was produced in the same manner as in Example 1. The ultrafiltration performance of the filter was 107 mL/min/200 mmHg. The filter was used as the above-mentioned filter for concentration, and the ascites concentration operation was performed at a flow rate of 70 mL per minute while the sieve coefficient of the simulated ascites reached 0.07, and then the ascites concentration operation was performed at an inflow rate of 70 mL per minute. The concentration ratio was 5 times or more, and the concentration time was 54 minutes, but the filtrate protein concentration was 0.210 g/dL.

[Comparative Example 7]

A polypene concentration filter was produced in the same manner as in Example 1. The ultrafiltration performance of the filter is 160 mL/min/200 mmHg. This filter was used as the above-mentioned filter for concentration, and the ascites concentration operation was performed at a flow rate of 120 mL per minute during the period of 1/4 of the simulated ascites, and then the ascites concentration operation was performed at an inflow rate of 120 mL per minute. When the concentration ratio was 5 times or more, the concentration time was 29.5 minutes, but the protein concentration in the filtrate became 1.753 g/dL.

[Comparative Example 8]

A polypene concentration filter was produced in the same manner as in Example 1. The ultrafiltration performance of the filter is 80 mL/min/200 mmHg. The filter was used as the above-mentioned filter for concentration, and the ascites concentration operation was performed at a flow rate of 120 mL per minute during the period of 1/4 of the simulated ascites, and the ascites concentration operation was performed at an inflow rate of 120 mL per minute. The water is treated, and the amount of the recovered liquid is measured. As a result, the target magnification is not reached, and it is necessary to rearrange the line and the rotary pump to perform additional concentration of the recovered liquid in the recovery container.

[Comparative Example 9]

A polypene concentration filter was produced in the same manner as in Example 2. The ultrafiltration performance of the filter is 110 mL/min/200 mmHg. This filter was used as the above-mentioned filter for concentration, and bovine plasma adjusted to a total protein concentration of 7 g/dL was used as simulated ascites, and 3 L was prepared. At the time point of processing 1/10 of the simulated ascites, the sieve coefficient reached 0.03, so before that, 70 mL per minute was used, and then the ascites concentration operation was performed at an inflow speed of 120 mL per minute. As a result, all the simulated ascites were treated and measured. When the amount of liquid is recovered, the result does not reach the target magnification, and it is necessary to rearrange the line and the rotary pump to perform additional concentration of the recovered liquid in the recovery container.

[Comparative Example 10]

A polypene concentration filter was produced in the same manner as in Example 2. The ultrafiltration performance of the filter is 110 mL/min/200 mmHg. The filter is used as the concentration filter, and the ascites concentration operation is performed at a flow rate of 120 mL per minute during the period of 1/4 of the simulated ascites, and the concentration of the ascites is performed at an inflow rate of 120 mL per minute. The concentration ratio was 5 times 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 filter for concentration was produced in the same manner as in Example 2. The ultrafiltration performance of the filter is 110 mL/min/200 mmHg. The filter was used as the concentration filter, and the ascites concentration operation was performed at a flow rate of 120 mL per minute while the sieve coefficient of the simulated ascites reached 0.04, and then the ascites concentration operation was performed at an inflow rate of 120 mL per minute. The concentration ratio was 5 times or more, and the concentration time was 30.4 minutes, but the filtrate protein concentration became 0.113 g/dL.

The measurement conditions and measurement results of Examples 1 to 12 and Comparative Examples 1 to 11 are shown in Table 1 and Table 2.

In Examples 1 to 12, the treatment time can be suppressed to within 57 minutes while controlling the protein concentration sent from the filtration side outlet of the ascites-concentrating filter to 100 mg/dL or less. On the other hand, in Comparative Example 1 and 2 in which the ascites inflow rate was 100 mL/min and 200 mL/min, and the total treatment time was controlled to a fixed value, and in Comparative Example 10 in which the inflow speed in the first step was set relatively, the filtration for concentration was used. The concentration on the filtrate side of the device was 100 mg/dL or more.

In the third example, the flow rate of the first stage was controlled to 30 mL per minute, and the flow rate of the second stage was controlled to 50 mL per minute. The filtrate protein concentration was the same as that of Examples 1 to 3. Degree, but the processing time becomes 70 minutes, exceeding 60 minutes of the time limit. Further, in Comparative Examples 4 and 8 in which a filter having a low ultrafiltration performance was used, concentration was not achieved five times or more in a single concentration, and additional concentration was necessary. Further, in Comparative Examples 5 and 7 in which a filter having a very high ultrafiltration performance was used, the protein was not sufficiently concentrated, and only a protein solution having a lower protein concentration was obtained.

Further, in Comparative Example 6 in which the time point of switching to the second stage was set to the time point of the sieve coefficient of 0.07, and the comparative example 11 in which the time point of switching to the second stage was set to the time point of the sieve coefficient of 0.04, the filtrate was used. The protein concentration on the side is 100 mg/dL or more. In Comparative Example 9 in which the protein concentration of ascites was set higher, concentration was not achieved five times or more, and concentration was necessary.

[Industrial availability]

According to the present invention, there can be provided a method and a manufacturing apparatus for producing a high-concentration protein solution which is a method for obtaining a concentrated protein solution by concentrating a thin protein solution such as ascites without causing clogging. The processing speed is lowered, and a thick protein solution having a high protein concentration can be obtained without burdening the implementer such as an additional concentration step.

1‧‧‧ storage container

1b‧‧‧Export (connection between storage container and first flow path)

2‧‧‧Recycling container

2a‧‧‧ entrance (connection between the recovery container and the fourth flow path)

3‧‧‧Filter filter

3a‧‧‧Inlet filter filter

3b‧‧‧Export of filter filter

3c‧‧‧ filtrate outlet (filter side outlet for ascites filtration)

4‧‧‧Concentration filter

4a‧‧‧ ascites inlet (inlet of ascites concentrate filter)

4b‧‧‧ Concentrate outlet (export of ascites concentrate filter)

4c‧‧‧ filtrate discharge port (filter side outlet for filter for ascites concentration)

5‧‧‧ pump (control agency)

14.15‧‧‧Control Department (Control Agency)

31‧‧‧1st flow path

32‧‧‧2nd flow path

33‧‧‧3rd flow path

34‧‧‧4th flow path

35‧‧‧5th flow path

41‧‧‧Control device (control mechanism)

100‧‧‧ Ascites filtration and concentration device

Claims (16)

A method for producing a high-concentration protein solution, comprising: a first step of allowing a low-concentration protein solution to pass through a line and having an ultrafiltration performance of 85 mL to 150 mL/min/200 mmHg and imparting it to a storage container storing a low-concentration protein solution a polyether-based hollow fiber membrane type ascites-concentrating filter for transporting a filtrate from a filtration side outlet of the filter, and delivering a high-concentration protein solution from the outlet of the filter; The high-concentration protein solution sent from the outlet of the filter is recovered in a recovery container, and the first step includes a first step of passing the low-concentration protein solution through the ascites-concentrating filter at a first flow rate, and second In the step, the low-concentration protein solution is passed through the ascites-concentrating filter at a second flow rate faster than the first flow rate at a time when the total amount of the low-concentration protein solution is supplied to the liquid amount by a predetermined amount or more. The method of claim 1, further comprising: setting a membrane area of the filter to A (m ² ), and setting the weight of the low-concentration protein solution treated in the first stage to V1 (kg) The protein concentration of the low-concentration protein solution is C (g/dL), the flow rate of the first stage is Qb1 (mL/min), and the filtration flow rate of the first stage is Qf1 (mL/ Min), when the ultrafiltration performance of the above filter is F (mL/min/200 mmHg), the second stage is switched to the above stage (1): 103.7≦-37log (A/V1) +log(Qb1/V1)+57log(F)-log(1/C)-log(Qb1/Qf1)≦112.6 (1). The method of claim 1 or 2, wherein the switching from the first flow rate to the second flow rate is It is carried out at a time point when at least 1/4 or more of the total amount of the low-concentration protein solution is supplied. The method of claim 1 or 2, wherein the first flow rate is 70 mL/min or less, and the second flow rate is 120 mL/min or less. The method of claim 1 or 2, wherein the first flow rate is 50 mL/min or less, and the second flow rate is 70 mL/min or less. The method of claim 1 or 2, wherein a total value of the first flow rate and the second flow rate is 100 mL/min or more, and a difference in flow rate between the first flow rate and the second flow rate is at least 20 mL/min or more. The method of claim 1 or 2, wherein in the first step, the protein concentration of the low-concentration protein solution is 5 g/dL or less. The method of claim 1 or 2, wherein in the first step, the protein concentration of the low-concentration protein solution is 3 g/dL or less. The method of claim 1 or 2, wherein, in the first step, when the sieve coefficient of the protein in the high-concentration protein solution sent from the filtration side outlet of the ascites-concentrating filter is a specific value or less, the first step is started. 2 stages. The method of claim 1 or 2, wherein, in the first step, when the sieve coefficient of the protein in the high-concentration protein solution sent from the filtration side outlet of the ascites-concentrating filter is at least 0.03 or less, the first step is started. 2 stages. The method of claim 1 or 2, 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 of claim 1 or 2, wherein in the first step, the protein concentration of the high-concentration protein solution sent from the outlet of the ascites-concentrating filter is 7 g/dL or more. The method of claim 1 or 2, wherein the line comprises a filter for ascites filtration. The method of claim 1 or 2, wherein the first step is performed by performing a first step of supplying a liquid to the ascites concentration filter at a first flow rate, and monitoring the inlet of the ascites concentration filter The protein concentration in the solution with the outlet is subjected to the second stage at a time when the sieve coefficient of the above protein falls to at least 0.03. The method according to claim 1 or 2, wherein the first step is controlled by: performing a first stage of supplying the liquid to the ascites concentration filter at a first flow rate, and monitoring the weight of the low-concentration protein solution; The second stage is carried out at a time point when the total amount of the low-concentration protein solution is more than a predetermined amount. A manufacturing apparatus for a high-concentration protein solution, comprising: a part comprising the first step, wherein the low-concentration protein solution passes through a line and has an ultrafiltration performance of 85 mL from a storage container in which a low-concentration protein solution is stored. 150mL/min/200mmHg and a hydrophilic polymer high-concentration hollow fiber membrane type ascites concentration filter, the filtrate is sent out from the filter side outlet of the filter, and a high-concentration protein solution is sent from the outlet of the filter; Included in the second step, the second step is to recover the high-concentration protein solution sent from the outlet of the filter into a recovery container, and the portion including the first step includes: a portion having a first stage, In the first stage, the low-concentration protein solution is passed through the ascites concentration filter at a first flow rate; and the second stage is a second stage, and the second stage is supplied from a total amount of the low-concentration protein solution to a specific amount or more. At a time point, the low-concentration protein solution is passed through the ascites concentration filter at a second flow rate faster than the first flow rate, and The membrane area of the filter is A (m ² ), and the weight of the low-concentration protein solution treated in the first step is V1 (kg), and the protein concentration of the low-concentration protein solution is set to C (g/dL), the flow rate of the first stage is set to Qb1 (mL/min), and the filtration flow rate of the first stage is set to Qf1 (mL/min), and the ultrafiltration performance of the filter is set to In the case of F (mL/min/200mmHg), when the following (1) is satisfied, the second stage is switched: 103.7≦-37log(A/V1)+log(Qb1/V1)+57log(F) -log(1/C)-log(Qb1/Qf1)≦112.6(1).