JPS6139826B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to an adsorbent for adsorbing blood proteins. Recently, plasma exchange therapy has been attracting attention due to its remarkable effectiveness in treating cancer diseases, autoimmune diseases, liver failure, and the like. This is thought to be to remove proteins such as albumin that are bound to antibodies, immunosuppressive factors, or metabolic products that cause the disease.
However, in plasma exchange therapy, the patient's plasma is discarded and replaced with healthy human plasma, which requires a large number of plasma each time, making treatment difficult for many people. Furthermore, it is a very wasteful therapy because it simultaneously removes unnecessary protein components and other useful components. On the other hand, purification therapy using an adsorbent removes only unnecessary components, so it has the advantage of requiring little or no replenishment solution.
No. 22178, JP-A-50-76219 and JP-A-51-
When activated carbon or porous resins such as those disclosed in No. 148291 are used, although low molecular weight substances such as creatinine and uric acid are adsorbed, almost no protein is adsorbed, or other substances other than the target protein are adsorbed. Since large amounts of other useful proteins, vitamins, sugars, etc. are also adsorbed at the same time, sufficient therapeutic effects for the above-mentioned diseases cannot be obtained. Also, JP-A-53-80381, 54-9183, 54-
Adsorbents made of organic polymers, such as those disclosed in No. 131586, had similar drawbacks and were not suitable for the purpose of the present invention. An object of the present invention is to provide an adsorbent for selectively adsorbing specific proteins in blood. Another object of the present invention is to provide an adsorbent useful for treating autoimmune diseases, cancer diseases, liver failure, etc. through blood purification. The adsorbent of the present invention has an average pore diameter of 30 to 3000 Å, and when the average pore diameter is D, the sum of the volumes of pores with pore diameters in the range of 0.8D to 1.2D is It is a porous material that occupies more than 80% of the total pore volume.
If the average pore diameter is smaller than 30 Å, hardly any protein will be adsorbed, and if the average pore diameter is 3000 Å or more, the adsorbent becomes brittle, which is inappropriate. The adsorbent of the present invention needs to have a sharp pore size distribution, and when the average pore diameter is D,
It is necessary that the sum of the volumes of pores with pore diameters in the range of 0.8D to 1.2D accounts for 80% or more of the total pore volume. If the pore size distribution is wider than this, the selective adsorption property will decrease and many types of proteins will be adsorbed at the same time, making it difficult to selectively adsorb only a specific protein, which is undesirable. .
Currently available activated carbon and organic porous resins do not have such a sharp pore size distribution and are therefore not included in the present invention. In the present invention, it is necessary to select the average pore diameter of the porous body depending on the molecular weight of the protein to be adsorbed. For example, when adsorbing proteins with a molecular weight of 500 to 20,000, using a porous material with an average pore diameter in the range of 30 to 150 Å will most selectively adsorb proteins with molecular weights in the above range. Also,
When adsorbing proteins with a molecular weight of 20,000 to 200,000, the average pore diameter is preferably 150 to 1,000 Å, and in the case of proteins with a molecular weight of 200,000 or more, it is 1,000 to 3,000 Å.
are preferably used. Specific examples of proteins to be adsorbed in the present invention include the following. First, proteins with a molecular weight of 500 to 20,000 include poisonous snakes, scorpions, poisonous sea urchins, poisonous spiders,
Toxic proteins secreted by frogs, bees, etc., lysozyme, cytochrome C, and immunoregulatory α-globulin, which is specifically found in the blood of cancer patients.
Examples include immunosuppressive factors called (IRA). As a protein with a molecular weight of 20,000 to 200,000, γ
- Globulin, albumin, and α ₁ -
Antitrypsin (α ₁ AT), C-Reactive protein
(CRP), _α1 -Acid glycoprotein (AAG),
Immunosuppressive acid protein (IAP), α-
Examples include protein-based immunosuppressive factors such as fetoprotein (AFP). γ-globulin is a protein with a molecular weight of approximately 160,000, and among these proteins, immunoglobulin is the main causative agent of autoimmune diseases. Diseases are treated by removing it from the blood, but in the case of γ-globulin,
Particularly when using a porous body having an average pore diameter of 350 to 900 Å (more preferably 400 to 700 Å),
Since adsorption is performed efficiently and selectively, γ
- When the purpose is to adsorb globulin, it is particularly preferable to use a porous body having an average pore diameter within the above range. The protein-based immunosuppressive factors mentioned above are found in the blood of cancer patients.
It has the effect of suppressing the immune system's attack on cancer cells. In addition, as a protein with a molecular weight of 200,000 or more, C ₁
Complements such as q, fibrinogen, microfibrin, antigen-antibody complexes (immune complexes), low density lipoproteins, etc. can be mentioned. Examples of the porous body used in the present invention include porous glass, porous silica, porous alumina, and porous porcelain. Porous glass is produced by melt-forming alkali borosilicate glass, followed by heat treatment in a transition temperature range, and then acid-treating fine phase-separated glass. Porous silica is produced by acid treatment of an aqueous sodium silicate solution. Further, porous alumina is produced by firing a hydrated alumina compact, and porous porcelain is produced by sintering a porcelain aggregate and a glass binder. Of course, as mentioned above, these porous bodies are
It has an average pore diameter of 3000 Å, and when the average pore diameter is D, the sum of the volumes of pores with pore diameters in the range of 0.8D to 1.2D accounts for 80% or more of the total pore volume. It is necessary to occupy. Among the above-mentioned porous materials, those having silanol groups on the surface are preferably used because they have high protein adsorption performance and hardly adsorb useful components such as sugars and vitamins. Therefore, among the above-mentioned materials, porous glass and porous silica are preferably used.
In particular, porous glass is most preferred because it has high adsorption performance and high mechanical strength. The pore volume of these porous bodies is preferably 0.1 cc/g or more and 2 c.c./g or less, particularly preferably 0.5 cc/g or more and 2 c.c./g or less. If it is less than 0.1 cc/g, the adsorption performance will not be sufficient, and if it is more than 2 c.c./g, the mechanical strength will be low. The porous body used in the present invention preferably has a particle size in the range of 4 to 270 mesh, especially for whole blood.
50 mesh, preferably 4 to 270 mesh for plasma and serum. The porous body used in the present invention may be used as is, but it is preferably used whose surface is coated with a hydrophilic polymer because it has good affinity with blood. Examples of hydrophilic polymers include:
Acrylic acid polymers, methacrylic acid polymers, acrylic ester polymers, methacrylic ester polymers, acrylamide polymers, polyvinyl alcohol polymers, polyvinylpyrrolidone,
Examples include cellulose nitrate and gelatin. Among these, acrylic ester polymers and methacrylic ester polymers are preferred. More particularly preferred are acrylic ester or methacrylic ester monomers represented by the following general formula () or () and polymerizable monomers having an epoxy group represented by the general formula (), () or (). It is a copolymer consisting of polymers. (However, in the above general formula, R ₁ , R ₁ ′, R ₁ ″ are hydrogen or a methyl group; R ₂ is a divalent alkylene group having 2 to 3 carbon atoms or poly(oxyalkylene) with or without a substituent. ) group; R ₃ is a divalent alkylene group having 1 to 3 carbon atoms or a poly(oxyalkylene) group with or without a substituent; R ₄ and R ₄ ' are hydrogen or an alkyl group having 1 to 3 carbon atoms; , the alkyl group may further have a hydroxyl group or an amino group) The porous body coated with the hydrophilic polymer of the present invention needs to be sterilized when used for therapeutic purposes, Steam sterilization is particularly preferred, but in order to prevent elution of the polymer from the coating layer, it contains a polymerizable monomer having an epoxy group represented by the above general formula (), (), () as a structural unit. It is recommended to use a copolymer as a coating material and to make it crosslinked and insolubilized by heat treatment or the like after coating.
The copolymerization ratio of the polymerizable monomer having an epoxy group is suitably 0.1 to 10% by weight. The method of coating the surface of a porous body with a hydrophilic polymer is to dissolve the hydrophilic polymer in an appropriate solvent such as methanol or ethanol, and coat the porous body by dipping, spraying, or wet coagulation. can. It is necessary that the coating layer imparts appropriate blood affinity to the porous body and does not significantly reduce the adsorption performance of the porous body. In this respect, the concentration of the hydrophilic polymer solution during the coating process is
A range of 0.05-2%, preferably 0.05-0.5% is suitable. When coated with a copolymer containing a monomer containing an epoxy group represented by the above general formula (), (), () as a constituent component, the temperature is 80 to 120℃.
Crosslinking and insolubilization can be achieved by heat treatment for 1 to 24 hours. The porous body used in the present invention can further enhance adsorption selectivity by introducing charges to the surface. If the protein to be adsorbed is a basic protein, a negatively charged group such as a carboxyl group or a sulfonic acid group is introduced; if the protein is an acidic protein, a positively charged group such as an amino group is introduced. do. The introduction of amino groups can be carried out by treating the porous material with an aminosilane compound such as γ-aminopropyltriethoxysilane, and the introduction of carboxyl groups can be carried out by reacting with succinic anhydride after aminosilanization, or by reacting with succinic anhydride or using carbodiimide. This can be carried out by reacting with succinic acid in the presence of succinic acid. Further, the sulfonic acid group can be introduced by, after the aminosilanization treatment, treating with glutaraldehyde in an acidic state to introduce an aldehyde group, and then treating this with taurine in an alkaline state. Furthermore, as another method for introducing charges, a method of coating the porous body with a polymer having a carboxyl group, a sulfonic acid group, or an amino group can be mentioned. Examples of such polymers include polyacrylic acid, polymethacrylic acid, polystyrene sulfonic acid, and copolymers of monomers and hydrophilic monomers that are constituent units of these polymers. The appropriate concentration of these polymers during coating is 0.05 to 5%, preferably 0.1 to 2%. For example, when albumin, which is an acidic protein, is to be adsorbed, albumin can be adsorbed with high selectivity by using a porous material having amino groups introduced onto its surface. The porous body used in the present invention needs to have a specific average pore diameter and pore diameter distribution as described above, and these values are values measured by a mercury porosimeter. Since the adsorbent of the present invention has a sharp pore size distribution, it has excellent selective adsorption properties for proteins. Therefore, by using a porous material with a specific average pore diameter depending on the molecular weight of the protein to be adsorbed, the target protein can be efficiently absorbed without adsorbing most of the useful components in the blood. Can be adsorbed. As mentioned above, the relationship between the molecular weight of a protein and the average pore diameter is
500 to 20,000, pore diameter 30 to 150 Å, molecular weight 2
It is 150 to 1000 Å for a molecular weight of 200,000 to 200,000, and 1000 to 3000 Å for a molecular weight of 200,000 or more. On the other hand, activated carbon and porous resins, which have traditionally been used to adsorb and remove harmful components in blood, have a wide pore size distribution, so they can absorb useful proteins, sugars, and vitamins other than the target protein. It also adsorbs other substances, which is not desirable. Hereinafter, the mode of use of the blood protein adsorbent of the present invention will be explained in more detail with reference to the drawings. FIG. 1 is an example of a column filled with the adsorbent of the present invention. The main body 1 has a blood inlet 2 and an outlet 3, and a filter 4 is provided at the inlet and outlet. Moreover, the adsorbent 5 of the present invention is filled between the filters. Blood enters through the inlet 2, passes through the filter 4, comes into contact with the adsorbent 5, and after proteins are adsorbed by the adsorbent, it exits the column through the outlet 3. When adsorbing and removing proteins in blood using the adsorbent of the present invention, as shown in FIG. A system that removes it and reintroduces it back into the body is usually available. In another system, as shown in Figure 3, blood is taken out of the body, blood cells and plasma are separated by a plasma separator 7, and only the plasma is passed through an adsorption column 1 to remove proteins by adsorption. One example is a system that mixes it with blood cell components again and returns it to the body.
These systems can be freely selected depending on the purpose. Moreover, it can also be used by being incorporated into a system other than such an extracorporeal circulation system. As described above, the adsorbent of the present invention can selectively adsorb and remove specific proteins in blood, making it possible to treat various diseases and disorders. Note that the term blood here includes not only whole blood but also plasma, serum, and the like. For example, in autoimmune diseases, antibodies (immunoglobulin, molecular weight approximately 16
10,000), which becomes the main causative agent of the disease. Antibodies exist in blood either alone or in the form of antigen-antibody complexes (immune complexes, molecular weight 200,000 to 1,000,000) in combination with antigens. Therefore, the average pore diameter is
Adsorb and remove immunoglobulins using adsorbents with a diameter of 350–900 Å, or with an average pore diameter of 1000–
Treatment can be performed by adsorbing and removing the antigen-antibody complex using a 3000 Å adsorbent. In addition, in organ transplantation, antibodies (immunoglobulin) against the organ are produced, causing a rejection reaction. However, by adsorbing and removing the antibodies using the above-mentioned adsorbent, rejection reactions can be suppressed. In addition, the blood of cancer patients contains immunoregulatory
α-globulin (IRA, molecular weight 500-10,000), α ₁ −
Antitrypsin (α ₁ AT), C-Reactive protein
(CRP, molecular weight approximately 140,000), α ₁ -Acid
glycoprotein (AAG), immunosuppressive
acid protein (IAP, molecular weight approximately 59,000), α-
Immunosuppressive factors considered to be antigens such as fetoprotein (AFP, molecular weight approximately 74,000) and antibodies (molecular weight 100,000 to 20,000
There are immunosuppressive factors that are thought to be
The immune system's attack on cancer cells is evaded.
The abundance of these immunosuppressive factors differs depending on the type of cancer, so the average pore diameter of the adsorbent is selected to remove the main components (30 to 150 Å for IRA; For others
150-1000 Å), cancer can be treated by adsorbing and removing immunosuppressive factors. In addition, in liver failure, metabolic products such as bilirubin are present in large quantities in the blood, bound to albumin.
Average pore diameter 150-1000Å as it shows jaundice symptoms
It can be treated by adsorbing and removing this using an adsorbent having an amino group on its surface. As described above, the present invention is useful for the treatment of various diseases and diseases, but if necessary, by using a column filled with two or more types of adsorbents or by using a plurality of columns, It is also possible to enhance the therapeutic effect or treat two or more diseases at the same time. The column of the present invention is usually sterilized before use, and preferred sterilization methods include high-pressure steam sterilization and gamma ray sterilization. Example 1 Porous glass CPG-10-75 manufactured by Electro Nucleonics (average pore diameter D = 90 Å,
The ratio of pore volume in 0.8D to 1.2D is 98%, the pore volume is 0.54 cc/g, the surface area is 174 m ² /g, and the particle size is 80 to 120 mesh) (Example 1-1). After introducing amino groups by refluxing propyltriethoxysilane, a carboxylated porous glass (Example 1-2) was obtained by reacting with succinic anhydride in dehydrated dioxane.
In addition, CPG-10-75 is immersed in a 1% ethanol solution of a copolymer consisting of 79% hydroxyethyl methacrylate, 20% methacrylic acid, and 1% glycidyl methacrylate, then dried and heat-treated to produce a hydrophilic hydrophilic material containing carboxyl groups. A porous glass coated with a polymer (Example 1-3) was obtained. Porous glass FPG manufactured by Wako Pure Chemical Industries, Ltd.
-100L (average pore diameter D = 96 Å, proportion of pore volume between 0.8D and 1.2D 82%, pore volume 0.60cc/g,
Example 1-5 was obtained by carboxylating Example 1-4 in the same manner ^as in Example 1-2. Example 1-6 was obtained by coating FPG-100L with a hydrophilic polymer in the same manner as in Example 1-3. In addition, porous glass CPG-10-170 manufactured by Electro Nucleonics (average pore diameter D =
220Å, 90% of the pore volume lies between 0.8D and 1.2D,
Pore volume 1.23cc/g, surface area ^140m2 /g, particle size 80~
120 mesh) in Example 1-7, CPG-10-240
(Average pore diameter D = 370 Å, proportion of pore volume between 0.8D and 1.2D 86%, pore volume 1.34cc/g, surface area
97m ² /g, particle size 80-120 mesh) in Example 1-8
And so. Also, for comparison, bead-shaped activated carbon
BAC-MU-L (manufactured by Kureha Chemical, wide pore size distribution)
was used (Example 1-9). The adsorption amounts of egg white lysozyme (manufactured by Sigma), cytochrome C (manufactured by Sigma, horse heart), and bovine serum albumin (manufactured by Sigma, Fraction V) were measured for these samples, and the results are shown in Table 1.

[Table] As shown in Table 1, the adsorbents of Examples 1-1 to 1-6 (average pore diameter of 150 Å or less) adsorb low molecular weight proteins such as lysozyme and cytochrome C well, but do not adsorb serum albumin at all. I didn't. In contrast, Examples 1-7 and 1-8 (average pore diameter
The porous material with a diameter of 150 Å or more had low selectivity between lysozyme and serum albumin. Furthermore, in Examples 1-9 in which activated carbon was used, the adsorption amounts of lysozyme and albumin were both low, and the selective adsorption properties were also poor. Example 2 Porous glass CPG-10-240 manufactured by Electro Nucleonics (see Example 1-8) (Example 2-1) was mixed with 79% by weight of hydroxyethyl methacrylate, 20% by weight of methacrylic acid, and glycidyl. A porous glass (Example 2-2) coated with a hydrophilic polymer was obtained by immersing a copolymer containing 1% by weight of methacrylate in a 1% by weight ethanol solution and then drying and heating the glass. Porous glass FPG manufactured by Wako Pure Chemical Industries, Ltd.
-250L (average pore diameter D = 220 Å, proportion of pore volume between 0.8D and 1.2D: 96%, pore volume 0.89cc/g,
Example 2-4 was obtained by coating Example ^2-3 with a hydrophilic polymer in the same manner as in Example 2-2. The company's porous glass FPG-100L (Example 1-
4) is referred to as Example 2-5, and this is referred to as Example 2-5.
Example 2-6 was obtained by coating with a hydrophilic polymer in the same manner as in Example 2-6. In addition, activated carbon beads
BAC-MU-L (see Examples 1-9) was used for comparison (Examples 2-7). Table 2 shows the adsorption amounts of bovine serum albumin (Fraction V, manufactured by Sigma) and bovine serum γ-globulin (Fraction, manufactured by Sigma). It is clear from the table that the porous bodies of Examples 2-1 to 2-4 (average pore diameter of 150 Å or more) were albumin, γ
- Sufficiently adsorbs globulin, but Example 2-
When a porous body with a diameter of 5,2-6 (average pore diameter of 150 Å or less) is used, albumin and γ-globulin are not adsorbed at all.

【table】

[Table] Example 3 Electro-Nucreonics porous glass CPG-10-240 (see Example 1-8) (Example 3-1), CPG-10-500 (average pore diameter D =
700 Å, 99% or more of pore volume between 0.8D and 1.2D, pore volume 0.87cc/g, surface area 39m ² /g, particle size 80
~120 meshes) (Example 3-2), CPG-10-
700 (average pore diameter D = 880 Å, proportion of pore volume between 0.8D and 1.2D is 99% or more, pore volume 1.25cc/
g, surface area 37 m ² /g, particle size 80-120 mesh) (Example 3-3) were taken, and 0.5 g each of bovine serum albumin (Fraction V, manufactured by Sigma) and bovine serum γ-globulin (manufactured by Sigma) were taken. 3 ml of a solution obtained by mixing and dissolving 2 g/dl of phosphate buffered saline in a phosphate buffered saline solution was added to each, and the mixture was shaken 120 times/min at 37°C.
The supernatant was sampled over time, albumin was quantified by the bromcresol green method, total protein was quantified by the Biuret method, and γ-globulin concentration was measured as the difference between total protein and albumin. Table 3 shows the pore diameter of the porous glass and the adsorption capacity for albumin and γ-globulin. Comparing this result with Example 2, it can be seen that a porous material with an average pore diameter of 350 to 900 Å has excellent selective adsorption of γ-globulin.

[Table] Example 4 Porous glass CPG-10-500 manufactured by Electro Nucleonics (see Example 3-2), CPG
-10-700 (see Example 3-3), and CPG-10
−1000 (average pore diameter D = 1300 Å, 0.8D ~ 1.2D
The proportion of pore volume in 90%, pore volume 1.05cc/
(25 g, surface area 28 m ² /g, particle size 80-120 mesh) were immersed in 200 ml of a 5% toluene solution of γ-aminopropyltriethoxysilane and heated under reflux overnight for amination treatment. After washing and drying the treated CPG with toluene, 10 g of it was taken, immersed in 100 ml of a 10% dioxane solution of succinic anhydride, and shaken at 40° C. for 7 hours to undergo carboxylation treatment. The treated CPG was washed with dioxane and dried. Take 0.5 g of the obtained carboxylated CPG (Examples 4-1, 4-2, and 4-3, respectively),
The adsorption properties of albumin and γ-globulin were examined in the same manner as in Example 3. The results are shown in Table 4.

[Table] Example 5 The carboxylated CPG-10-1000 having a pore size of 1300 Å prepared in Example 4 was coated with a hydroxyethyl methacrylate/methacrylic acid copolymer by a spray method (coverage rate 0.2%). When the adsorption properties of albumin and γ-globulin were measured using the same method as in Example 3, the adsorption capacity after 3 hours was 15.0 mg/g for albumin and 75.0 mg/g for γ-globulin. Therefore, this coating process involves carboxylation.
It can be seen that the albumin adsorption property of CPG is reduced and the γ-globulin adsorption property is increased. Example 6 Porous glass CPG-10-1000 manufactured by Electro Nucleonics (see Example 4) was coated with a methanol solution of 0.5% polyacrylic acid by a spray method (coverage rate 0.5%) and heated at 120°C. Heat treatment was performed for 2 hours. Regarding this, when the adsorption properties of albumin and γ-globulin were measured in the same manner as in Example 3,
The adsorption capacity after 3 hours was albumin 42.0 mg/g, γ
- Globulin was 60.0 mg/g. Therefore, it can be seen that the γ-globulin adsorption ability of CPG can also be increased by coating it with a polymer having a carboxyl group. Example 7 Porous glass CPG-10-500 manufactured by Electro Nucleonics (see Example 3-2) was subjected to aminosilanization treatment in the same manner as in Example 4, and 5 g of this was taken and treated with 1N of 5% glutaraldehyde. 50 ml of hydrochloric acid solution was added, and the mixture was shaken and reacted at room temperature for 17 hours. This CPG was thoroughly washed with water, and then 50 ml of a 1N sodium hydroxide solution containing 5% taurine was added and reacted by shaking at room temperature for 8 hours. Regarding the sulfonated CPG in which a sulfonic acid group was introduced at the terminal using this method, albumin and γ were prepared in the same manner as in Example 3.
- When the adsorption capacity of globulin was measured, the adsorption capacity after 3 hours was 25.8 mg/g for albumin and 63.6 mg/g for γ-globulin. Therefore, it can be seen that the selective adsorption of γ-globulin increases more with sulfonation than with carboxylation. Example 8 Porous glass CPG-10-500 manufactured by Electro Nucleonics (see Example 3-2), CPG
-10-700 (see Example 3-3), CPG-10-
Take 25g each of 1000 (see Example 4),
Each was immersed in 200 ml of a 5% toluene solution of γ-aminopropyltriethoxysilane and heated under reflux overnight for amination treatment (these were used in Examples 8-2, 8-4, and 8-6, respectively). Raw CPG
(Examples 8-1, 8-3, 8-5) and amination
CPG (Examples 8-2, 8-4, 8-6) respectively
0.5 g each was taken and the adsorption performance of albumin and γ-globulin was measured in the same manner as in Example 3. The results are shown in Table 5. It can be seen that for any pore size, the amination treatment increases the albumin adsorption capacity and decreases the globulin adsorption capacity.

【table】

[Table] Example 9 Porous glass CPG-10-1400 manufactured by Electro Nucleonics (average pore diameter D = 1400
Å, proportion of pore volume in 0.8D~1.2D 88%, pore volume 0.94cc/g, surface area ^18m2 /g, particle size 80~120
Mesh) was aminated by heating to reflux with γ-aminopropylethoxysilane in toluene. Next, by reacting with succinic anhydride in dehydrated dioxane, CPG-10-1400 (Example 9-1) having carboxyl groups introduced onto the surface was obtained. CPG-10-2000 manufactured by the company (average pore diameter D =
2800Å, 99% of the pore volume located between 0.8D and 1.2D
Above, pore volume 0.66 cc/g, surface area 8.3 m ² /g) was used as Example 9-2, and this was used as Example 9-1.
Example 9-3 was obtained by introducing a carboxyl group onto the surface in the same manner as in Example 9-3. Also, CPG−10−
1% by weight of a copolymer prepared from 2000 with 79% by weight of hydroxyethyl methacrylate, 20% by weight of methacrylic acid, and 1% by weight of chrycidyl methacrylate.
CPG coated with a hydrophilic polymer by immersing it in an ethanol solution and then drying and heating it.
10-2000 (Example 9-4) was obtained. FPG-2000L manufactured by Wako Pure Chemical Industries, Ltd. (average pore diameter D
= 2200Å, percentage of pore volume between 0.8D and 1.2D 90
%, pore volume 0.92 cc/g, surface area 14 m ² /g) was used as Example 9-5, and a carboxyl group was introduced onto the surface in the same manner as in Example 9-1. -6. CPG manufactured by Electro Nucleonics
10-700 (see Example 3-3) was used as Example 9-7, and a carboxyl group was introduced on the surface in the same manner as Example 9-1.
-8. For these porous materials, urease (molecular weight
470,000, type manufactured by Sigma) and bovine serum γ-globulin (molecular weight 160,000, fraction manufactured by Sigma)
We investigated the adsorption performance of Initial concentration of each protein is 2g/dl
The protein adsorption capacity was calculated by colorimetrically quantifying the concentration of the supernatant solution using a phosphate buffered saline solution, a bath ratio of 6 ml/1 g adsorbent, and shaking at 37° C. for 3 hours. The results are shown in Table 6.

[Table] As is clear from Table 6, the average pore diameter is 1000
In Examples 9-1 to 9-6, in which a porous material having a diameter in the range of ~3000 Å was used, urease, which is a high molecular weight protein, was selectively adsorbed, and γ-globulin, which has a low molecular weight, was hardly adsorbed. On the other hand, the average pore diameter
In Examples 9-7 and 9-8, in which the particle size was smaller than 1000 Å, hardly any urease was adsorbed, and a large amount of γ-globulin was adsorbed. Example 10 Porous glass CPG-10-1400 manufactured by Electro Nucleonics (see Example 9) was used as Example 10-1, and porous glass manufactured by Wako Pure Chemical Industries, Ltd.
FPG-700L (average pore diameter D = 720Å, 0.8D ~
Percentage of pore volume in 1.2D 99%, pore volume 0.95
cc/g, surface area 37m ² /g, particle size 80-120 mesh)
was used as Example 10-2, in which a carboxyl group was introduced into the surface in the same manner as in Example 9-1, and the adsorption performance of catalase (molecular weight 240,000, manufactured by Miles Laboratory, bovine liver) was investigated. The experimental conditions were the same as in Example 9 except that the initial protein concentration was 2.3 g/dl. The results are shown in Table 7. molecular weight
To adsorb 240,000 catalase, the average pore size
It can be seen that a porous material of 1000 Å or more is required.

[Table] Example 11 Porous glass CPG-10-350 manufactured by Electro Nucleonics (average pore diameter D = 380
Å, 86% of pore volume in 0.8D~1.2D, pore volume 1.39cc/g, surface area ^90m2 /g, particle size 80~120
Hydroxyethyl methacrylate
After soaking in a 0.25 wt% ethanol solution of a copolymer consisting of 99.5 wt% glycidyl methacrylate and 0.5 wt% glycidyl methacrylate, 50 c.c. Blood from a rabbit (male, 3.46 kg) was poured into a packed polypropylene column (with polyester 180 mesh filters at the column inlet and outlet) at a flow rate of approximately 5.
Extracorporeal circulation was performed at ml/min for 1 hour. Blood sampled from the arterial and venous sides was centrifuged to obtain plasma. This plasma was subjected to high-performance liquid chromatography (equipment ALC/GPC244 manufactured by Waters Associates Inc.).
Mold, column G-3000SW manufactured by Toyo Soda Co., Ltd. (inner diameter 7.5
mm, length 600 mm), eluent 1/15M phosphate buffer (contains 0.15M NaCl, PH6.0), flow rate 1.0ml/min, detector UV (280nm)). The removal rates of albumin and γ-globulin were determined. As shown in Table 8, approximately 70% of γ-globulin could be removed in 1 hour. During this time, albumin decreased by only 20%.

[Table] Example 12 Polypropylene column packed with 2g of porous glass CPG-10-75 manufactured by Electro Nucleonics (see Example 1) (see Example 11)
15 ml of rabbit blood containing 120 mg of lysozyme to 37
℃ and a flow rate of about 3 ml/min for 3 hours. Table 9 shows the results of time changes in the concentrations of lysozyme, albumin, and γ-globulin determined in the same manner as in Example 11. As is clear from the table, 100% of lysozyme could be removed in 3 hours, but albumin and γ-globulin did not decrease at all during this time.

[Table] Example 13 To 12 ml of rabbit anti-BSA antiserum obtained by immunizing a rabbit with bovine serum albumin (BSA), 100 mg of bovine serum albumin was added and reacted at 37°C for 30 minutes. Thereafter, it was passed through a 0.2μ filter to obtain rabbit serum containing soluble immune complexes. 10 ml of the rabbit serum was added to a polypropylene column (same column as in Example 12) packed with 2 g of carboxylated CPG-10-2000 (see Example 9-3) at 37°C for 3 hours at a flow rate of about 3 ml/min. It circulated. Changes in immune complex concentration were measured using high-performance liquid chromatography (the column was G-4000 manufactured by Toyo Soda Co., Ltd.).
It was analyzed under the same conditions as Example 11 except for SW (ID7.5mm, 600mm). The results are shown in Table 10, where 30% of immune complexes were removed within 3 hours, but other proteins (mostly albumin and γ-globulin) were reduced by only 7%. It was found that immune complexes could be selectively removed from serum.

【table】 [Brief explanation of the drawing]

FIG. 1 is a diagram showing an example of a column filled with the adsorbent of the present invention. FIGS. 2 and 3 are examples of a system for adsorbing and removing proteins in blood using the adsorbent of the present invention.

Claims (1)

[Claims] 1. Has an average pore diameter of 30 to 3000 Å, and where the average pore diameter is D, the pore diameter is 0.8D to
A blood protein adsorbent characterized by being a porous material in which the sum of the volumes of pores in the range of 1.2D accounts for 80% or more of the total pore volume. 2. The adsorbent according to claim 1, wherein the porous body has an average pore diameter in the range of 30 to 150 Å, and the protein has a molecular weight of 500 to 20,000. 3. The adsorbent according to claim 1, wherein the porous body has an average pore diameter in the range of 150 to 1000 Å, and the protein has a molecular weight of 20,000 to 200,000. 4. The adsorbent according to claim 1, wherein the porous body has an average pore diameter in the range of 1000 to 3000 Å, and the protein has a molecular weight of 200,000 or more. 5. The adsorbent according to claim 1, wherein the porous body has an average diameter in the range of 350 to 900 Å, and the protein is γ-globulin. 6. The adsorbent according to any one of claims 1 to 5, wherein the porous body is a porous body whose surface is coated with a hydrophilic polymer. 7 The hydrophilic polymer is an acrylic acid polymer, a methacrylic acid polymer, an acrylic ester polymer, a methacrylic ester polymer, an acrylamide polymer, a polyvinyl alcohol polymer,
The adsorbent according to claim 6, which is a polymer selected from the group consisting of polyvinylpyrrolidone, cellulose nitrate, and gelatin. 8. The adsorbent according to claim 6, wherein the hydrophilic polymer is an acrylic ester polymer or a methacrylic ester polymer. 9. The adsorbent according to claim 6, wherein the hydrophilic polymer is an acrylic ester polymer or a methacrylic ester polymer copolymerized with a polymerizable monomer having an epoxy group. 10. The adsorbent according to any one of claims 1 to 5, wherein the porous body has a negative charge on its surface. 11. The adsorbent according to any one of claims 1 to 5, wherein the porous body has a carboxyl group or a sulfonic acid group on its surface. 12. The adsorbent according to any one of claims 1 to 5, wherein the porous body has a silanol group on its surface. 13. The adsorbent according to any one of claims 1 to 5, wherein the porous body is a porous glass obtained by acid-treating alkali borosilicate glass having a fine phase separation structure. . 14 Claims 1 to 5, wherein the porous body is a porous body having a pore volume in the range of 0.5 to 2 c.c./g.
The adsorbent described in any of the paragraphs.