JPH01181875A - Adsorptive body of immune complex and removing device for immune complex with it - Google Patents

Abstract

PURPOSE:To selectively adsorb approximately an immune complex only without losing almost the effective component in a body fluid by obtaining the adsorbent of the immune complex composed by fixing the compound having an anion functional group at a water insoluble porous support. CONSTITUTION:The preferably elimination limit molecule quantity of a water insoluble porous support is desirable at 400,000 or above and 60,000,000 or below. Next, concerning the porous construction of the water insoluble porous support, the total porosity is more preferable than the surface porosity and since the hole volume is large than the adsorption capacity, it is desirable that it is 20% or above. As the compound having an anion functional group, it may be the compound having one anion functional group per polymer or a polyamine compound having plural anion functional groups. The adsorbent is made into the condition in which the compound having the anion functional group is fixed at the water insoluble porous support.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to an adsorbent for adsorbing and removing harmful components from body fluids and a removal device using the adsorbent. More specifically, the present invention relates to an adsorbent for removing immune complexes from body fluids and suppressing autoimmune diseases, and an immune complex removal device using the adsorbent.

[Problem 8 to be solved by conventional techniques and inventions] There are numerous types of infectious foreign substances such as viruses, bacteria, molds, and parasites around us.

All of these can cause diseases in living organisms, and if they multiply unrestricted within the organism, they will eventually kill the organism. Therefore, the body has an immune system as a function to protect against this. Antibody molecules play a central role in this immune system. In other words, when a foreign substance such as a virus or bacteria acts as an antigen and stimulates the immune system, antibody molecules that selectively bind to the antigen are produced.

On the other hand, in autoimmune diseases, a humoral or cellular immune response occurs against antigens of one's own cells and tissues rather than foreign organisms, and antibodies against oneself (hereinafter referred to as autoantibodies) and antigen-antibodies are produced in a specific manner. A complex formed by reacting with each other (
A large amount of immune complexes (hereinafter referred to as immune complexes) are generated. Immune complexes in body fluids are not processed by physiological elimination mechanisms and are deposited in tissues such as renal glomeruli, joint synovium, lungs, and blood vessel walls, directly contributing to the appearance of pathological conditions characteristic of autoimmune diseases. There are often conflicts.

Systemic lupus erythematosus (
In SLE (hereinafter referred to as SLE), antibodies (hereinafter referred to as anti-DNA antibodies) against nuclear components of cells, particularly deoxyribonucleic acid (hereinafter referred to as DNA), appear in body fluids, and these autoantibodies themselves bind to antigens and form immune complexes. A mechanism has been proposed that causes tissue damage by forming and depositing in tissues. In the case of SLH, the generated anti-DNA antibodies form immune complexes with cell-derived DNA that also leaked into the bloodstream.
It is known that vasculitis and lupus nephritis occur when deposited on blood vessel walls and renal glomerular basement membranes, respectively.
In fact, many cases of SLE result in death due to renal failure.

Immune complexes between the autoantibodies and the corresponding antigens produced in this way cause various symptoms, so controlling immune complexes is extremely important for treatment.

Steroids, immunosuppressants, immunomodulators, anti-inflammatory agents, and the like have been widely used in the treatment of SLH for the purpose of suppressing the production of immune complexes.

Among these, steroids are the most commonly used, and short-term ultra-dose therapy called pulse therapy is also often performed. However, since steroids tend to cause side effects even when administered in small doses, it is obvious that short-term ultra-dose therapy with steroids tends to cause even greater side effects. In addition, these drugs are often used for a long period of time, in which case side effects are more likely to occur, and the dosage must be gradually increased due to drug resistance, so in some cases, these drugs may not be used. In many cases, it is impossible or does not have sufficient effect. In particular, although suppression of anti-DNA antibodies and immune complexes is most necessary during the active phase of SLH, strong therapy using drugs such as pulse therapy and immunosuppressants cannot be used for the reasons mentioned above. There are also many.

On the other hand, as an approach different from these drug therapies, attempts have been made to directly remove immune complexes from body fluids by extracorporeal circulation.

The simplest method is so-called plasma exchange therapy, in which the patient's plasma containing immune complexes is exchanged with the plasma of a healthy person. This method significantly reduces immune complexes in the blood;
Symptoms are showing improvement. However, this method not only requires a large amount of healthy plasma and is expensive;
Because of the risk of infection such as serum hepatitis during the therapeutic treatment, it has not become widely used.

In plasma exchange therapy, all components in plasma are removed and replaced with healthy plasma.In contrast, in order to selectively remove immune complexes, which are pathogenic substances, the pathogenic substances are separated by molecular size. A plasma separation membrane method has been developed to separate In this method, plasma is separated into a high molecular weight fraction and a low molecular weight fraction using a membrane, the high molecular weight fraction containing pathogenic substances is discarded, and the low molecular weight fraction containing the main protein albumin is discarded. When returned to the patient, the immune complex has a molecular weight of approximately 18
Because the main component is a complex of IgG (immunoglobulin G) and antigen, and its molecular weight distribution is wide, separation from albumin, normal IgG, and IgM (immunoglobulin M) is not always sufficient, and immune complex It also has the disadvantage that a large amount of these substances are removed when the body is removed, and all proteins with a molecular weight equal to or higher than that of the pathogenic substance are removed.

Therefore, it has been desired to develop a means for removing immune complexes, which are pathogenic substances, more selectively, with almost no loss of useful components in body fluids.

As a result of extensive research in view of the above circumstances, the present inventors have discovered an adsorbent that can selectively adsorb almost only immune complexes without losing most of the active ingredients in body fluids, and have completed the present invention. It's arrived.

[Means to solve the problem]

That is, the present invention provides an adsorbent for an immune complex in which a compound having an anionic functional group is immobilized on a water-insoluble porous carrier, a container having an inlet and an outlet for a fluid, and a fluid and components contained in the fluid. The present invention relates to an immune complex removal device comprising a filter through which the immune complex adsorbent can pass through but not the immune complex adsorbent, and an immune complex removal device comprising the immune complex adsorbent filled in the container.

[Example] In this specification, body fluids include blood, plasma, serum, ascites, lymph fluid, intraarticular fluid, and fractionated components obtained therefrom.
and other biologically derived humoral components.

In the present invention, the water-insoluble porous carrier refers to a substance that is used to immobilize a compound having an anionic functional group and has the property of being insoluble in water. The water-insoluble porous carrier used in the present invention preferably has large, continuous pores. In other words, immune complexes are mainly composed of IgG and antigens, and have a molecular weight of 180,000 or more, and it is expected that there are large molecules with a molecular weight of around 1 million. It is necessary that the composite easily penetrate into the porous body.

There are various methods for measuring the pore diameter, and the mercury intrusion method is the most commonly used, but it is difficult to apply to hydrophilic porous materials. Exclusion limit molecular weight is often used as an alternative guideline for pore diameter, and can be applied to both hydrophilic porous materials and hydrophobic porous materials. The exclusion limit molecular weight is the bottom (
For example, as described in Hiroyuki Hatano, Toshihiko Hana, Experimental High Performance Liquid Chromatography, Kagaku Doujin), among the molecules that cannot enter (excluded) the pores in gel permeation chromatography, molecules with the smallest molecular weight The molecular weight of

It is known that the exclusion limit molecular weight varies depending on the target compound, and in general, globular proteins, dextran, polyethylene glycol, etc. have been well investigated, but in the case of the carrier used in the present invention, the molecular weight that is most similar to the immune complex is It is appropriate to use the values obtained using globular proteins that are thought to be present.

As a result of studies using various water-insoluble porous carriers with different exclusion limits, it was revealed that the range of pore diameters suitable for adsorption of immune complexes is from 400,000 to 80,000,000 to 80,000,000. That is, when a water-insoluble porous carrier having an exclusion limit molecular weight of less than 400,000 is used, the amount of immune complex adsorbed is too small to be practical. On the other hand, as the exclusion limit molecular weight becomes thicker, the adsorption amount of the immune complex increases but eventually reaches a plateau.When the exclusion limit molecular weight exceeds 60 million, the surface area is too small and the adsorption amount not only decreases noticeably, but also Adsorption of substances other than the desired immune complex, that is, non-specific adsorption, increases and selectivity decreases significantly.

Therefore, the preferred exclusion limit molecular weight of the water-insoluble porous carrier used in the present invention is 400,000 to 80 million,
More preferably, 8
It is preferable that the number is 00,000 or more and 20,000,000 or less.

Next, regarding the porous structure of the water-insoluble porous carrier, total porosity is preferable to surface porosity, and the pore volume is desirably 20% or more from the viewpoint of high adsorption capacity.

The shape of the water-insoluble porous carrier can be selected from any shape such as granules, spheres, fibers, membranes, and hollow fibers. When using a granular water-insoluble porous carrier, if the particle size is less than 1 mm, the pressure loss will be large;
If it exceeds 0m+, the adsorption capacity is small, so it is preferable that the adsorption capacity is 1 or more and 5000 or less.

The water-insoluble porous carrier used in the present invention may be either organic or inorganic, but it is preferably one that has little adsorption (so-called non-specific adsorption) of body fluid components other than the target immune complex. It is preferable that the water-insoluble porous carrier is hydrophilic rather than hydrophobic, since non-specific adsorption is less likely to occur if the carrier is hydrophilic.

Furthermore, it is advantageous if a functional group that can be used for a ligand immobilization reaction is present on the surface of the water-insoluble porous carrier. Typical examples of these functional groups include hydroxyl group, amino group, aldehyde group, alkyl group, thiol group, silanol group, amide group, epoxy group, halogen group, succinylimide group, acid anhydride group, etc. It is not limited to these. Further, it is particularly preferable that the water-insoluble porous carrier is made of a compound having a hydroxyl group among the above-mentioned functional groups, since non-specific adsorption is small. Needless to say, water-insoluble porous carriers into which these functional groups are introduced as spacers can also be used.

Representative examples of water-insoluble porous carriers used in the present invention include:
Soft porous materials such as agarose, dextran, and polyacrylamide, inorganic porous materials such as porous glass and porous silica gel, synthetic polymers such as polymethyl methacrylate, polyvinyl alcohol, styrene, and divinylbenzene copolymers, and/or cellulose. Examples include, but are not limited to, porous polymer hard gels made from natural polymers such as .

When using the adsorbent of the present invention for extracorporeal circulation treatment, blood,
Due to the need to flow high viscosity fluids such as blood plasma at high speeds, it is preferred to use a rigid, water-insoluble porous carrier that has sufficient mechanical strength to avoid compaction. In other words, the hard water-insoluble porous carrier is as shown in the reference example below.
When a water-insoluble porous carrier is uniformly packed into a cylindrical column and an aqueous fluid is passed through the column, the relationship between pressure loss and flow rate is linear up to at least 0.3 kg/c.

As the anionic functional group of the adsorbent of the present invention, any functional group can be used as long as it is negatively charged near neutral pH. Typical examples of these include carboxyl groups,
Examples include, but are not limited to, sulfonic acid groups, sulfone groups, sulfuric acid ester groups, silanol groups, phosphoric acid ester groups, and phenolic hydroxyl groups.

The compound having an anionic functional group may be a compound having one anionic functional group per molecule, or a polyanionic compound having a plurality of anionic functional groups. Polyanionic compounds are preferred because they have a high affinity for immune complexes and can easily introduce many anionic functional groups into a unit amount of water-insoluble porous carrier. Among these, polyanionic compounds having a molecular weight of 1000 or more are preferred in terms of affinity and the amount of anionic functional group introduced. The anionic functional group that the polyanion compound has is 1
It may be a type or a plurality of types.

Representative examples of polyanionic compounds used in the present invention include:
Synthetic polyanionic compounds such as polyacrylic acid, polyvinyl sulfuric acid, polyvinyl sulfonic acid, polyvinyl phosphoric acid, polystyrene sulfonic acid, polystyrene phosphoric acid, polyglutamic acid, polyaspartic acid, polymethacrylic acid, polyphosphoric acid, styrene-maleic acid copolymer, and anionic functional group-containing polysaccharides such as heparin, dextran sulfate, chondroitin, chondroitin sulfate, phosphomannan, chitin, and chitosan, but are not limited thereto.

The number of compounds having anionic functional groups immobilized on the adsorbent of the present invention may be one type, or two or more types.

The adsorbent of the present invention is one in which a compound having an anionic functional group is immobilized on a water-insoluble porous carrier. In order to obtain a state in which such a compound having an anionic functional group is immobilized, various known methods can be used without any particular limitations to introduce the anionic functional group into the water-insoluble porous carrier. However, typical methods for introducing an anionic functional group into a carrier to obtain a state in which a compound having such an anionic functional group is immobilized are as follows: (1) Anionic functional groups or easily anionic A method of forming an adsorbent by polymerization using a compound containing a functional group that can be converted into a functional group as a monomer or a crosslinking agent, (a method of immobilizing a compound containing a dianionic functional group on a water-insoluble porous carrier, G) A method of immobilizing a compound having an anionic functional group on a water-insoluble porous carrier by directly reacting the compound forming the anionic functional group with the water-insoluble porous carrier.

Of course, an anionic functional group-containing compound such as glass, silica, alumina, etc. that originally contains an anionic functional group may be used as the adsorbent.

Typical examples of monomers or crosslinking agents containing anionic functional groups or functional groups that can be easily converted into anionic functional groups used in method (1) include acrylic acid and its esters, methacrylic acid and its esters, and styrene. Examples include, but are not limited to, sulfonic acids.

(Method 2, that is, a method for immobilizing a compound containing an anionic functional group on a water-insoluble porous carrier, includes methods such as physical adsorption, ionic bonding, and covalent bonding. However, since it is important for the storage stability and stability of the adsorbent that the anionic functional group-containing compound not be desorbed, a covalent bonding method that allows for strong immobilization is preferable.

- When an anionic functional group-containing compound is immobilized by a covalent bond, it is preferable that the anionic functional group-containing compound has a functional group other than the anionic functional group that can be used for immobilization.

Typical examples of functional groups that can be used for immobilization include amino groups,
Examples include, but are not limited to, amide groups, carboxyl groups, acid anhydride groups, succinylimide groups, hydroxyl groups, thiol groups, aldehyde groups, halogen groups, epoxy groups, and silanol groups.

There are many anionic functional group-containing compounds having these functional groups, but sulfanilic acid,
Examples include phosphorylethanolamine, glycine, and taurine.

Among compounds containing anionic functional groups, typical examples of compounds containing sulfate groups include sulfate esters of hydroxyl group-containing compounds such as alcohols, sugars, and glycols. Partial sulfate ester compounds, especially sulfate esters of sugars, contain both sulfate groups and functional groups necessary for immobilization, and have high biocompatibility and activity.
Further, sulfated polysaccharides are particularly preferred because they are easily defined in water-insoluble porous carriers.

Next, by the method (3), that is, by reacting the compound forming an anionic functional group with the water-insoluble porous carrier, the compound having an anionic functional group is immobilized on the water-insoluble porous carrier, and the anionic functional group is immobilized on the water-insoluble porous carrier. A typical example of a method for introducing a functional group is a reaction in which a sulfate ester group is introduced into a hydroxyl group-containing porous carrier. In this case, the sulfate ester group can be directly introduced by reacting the hydroxyl group-containing water-insoluble porous carrier with a reagent such as chlorosulfonic acid or concentrated sulfuric acid.

The amount of anionic functional group introduced is 0.000000000000000000 per ml adsorbent. It is preferably 01μ■o1 or more and 10m■O1 or less.

0. If it is less than O1μs+o1, the adsorption capacity is insufficient, and if it exceeds 1011o1, there will be too much non-specific adsorption, making it difficult to put it to practical use. A more preferable amount of anionic functional group to be introduced is 1 .mu.O1 or more and 100 .mu.mol or less.

There are various ways in which the adsorbent of the present invention can be used therapeutically.

The simplest method is to draw the patient's blood outside the body and store it in a blood bag, mix it with the adsorbent of the present invention to remove immune complexes, then pass it through a filter to remove the adsorbent, and then transfer the blood to the patient. There is a way to get it back. Although this method does not require complicated equipment, it has the drawbacks that the amount of treatment per treatment is small, the treatment takes time, and the operation is complicated.

In the next method, the adsorbent is packed into a column and installed in an extracorporeal circulation circuit, and adsorption and removal is performed online. In other words, a container having an inlet and an outlet for fluid can pass through the fluid and the components contained in the fluid, but an immune complex adsorbent consisting of a water-insoluble porous carrier with a compound having an anionic functional group immobilized thereon can pass through the container. A simple and preferred method is to pass the body fluid through an immune complex removal device comprising an impermeable filter and an adsorbent for the immune complex filled in the container.

FIG. 2 shows a schematic cross-sectional view of an embodiment of the immune complex removal device of the present invention. In Figure 2, (1) and (2) are the inlet and outlet of the respective fluids, (3) is the adsorbent of the present invention, (4) and (S are the fluids and the components contained in the fluids can pass through. However, the adsorbent of the present invention cannot pass through a filter or mesh, (6) is a column, and (6) is a container.

Here, the filter on the fluid inlet side may not be present.

Hereinafter, the present invention will be explained in more detail with reference to Examples.
The present invention is not limited to such embodiments.

Reference Example: Agarose gel (B from Blorado) was placed on a glass cylindrical column (inner diameter 91 m, column length 150 + u) equipped with filters with a pore size of 15 IxR at both ends.
logel Asm, particle size 50-100 mesh), polymer hard gel (Toyo Pearl Awe manufactured by Toyo Soda Kogyo ■)
s, particle size 50-100 tm, and Cellulofine GC700 manufactured by Chisso ■, particle size 45-105. ca) was uniformly filled in each column, water was passed through the column using a beristaltic pump, and the relationship between the flow rate and pressure loss ΔP was determined. The results are shown in FIG.

As is clear from the figure, agarose gel, which is a soft gel, becomes compacted when the flow rate exceeds a certain level, and the flow rate does not increase even if the pressure increases, whereas hard gels such as Toyopearl and Cellulofine do not increase pressure. The flow rate increases approximately in proportion to.

Production Example 1 Porous cellulose gel CK Gel AS (trade name, manufactured by Chisso ■, exclusion limit molecular weight for globular proteins 50 million, particle size 45-105, ca) 20% Na in 100 ml
40 g of OH 120 g of heptane and the nonionic surfactant Tween 20 (trade name, manufactured by Kao Atlas ■)
Added 0 drops. After stirring at 40° C. for 2 hours, 50 g of epichlorohydrin was added and stirred for 2 hours, and the gel was washed and filtered with water to obtain a cellulose gel into which epoxy groups were introduced (hereinafter referred to as epoxidized gel).

Example 1 5 ml of the epoxidized gel obtained in Production Example 1 was added with 0.0 ml of sulfanilic acid. A solution of 1 g of L' dissolved in 10 ml of water and adjusted to pH 9.9 was added, shaken at room temperature for 24 hours, and 0.5% monoethanolamine aqueous solution was added and shaken to seal unreacted epoxy groups. A cellulose gel with immobilized sulfanilic acid was obtained. The amount of anionic functional groups introduced by immobilized sulfanilic acid is 6.5μ per ml of adsorbent.
It was mol.

Example 2 In 5 ml of the epoxidized gel obtained in Production Example 1, 0.1 g of phosphorylethanolamine was dissolved in 10 ml of water.
Add the solution adjusted to H9,8, shake at 40°C for 4 hours, add 0.5% monoethanolamine aqueous solution and shake to seal unreacted epoxy groups to form a cellulose gel with phosphorylethanolamine immobilized. I got it. The amount of anionic functional groups introduced by the immobilized phosphorylethanolamine was 4 μι/ml of adsorbent.

Example 3 5 ml of the epoxidized gel obtained in Production Example 1 had a molecular weight of about 500.
0, dextran sodium sulfate with 15% sulfur content 4
Add g and 5 ml of water to adjust the pH to 9 and incubate at 45°C for 16 hours.
Shake for hours. Thereafter, the gel was filtered, washed in this order with 2M saline solution, 0.5M saline solution, and water, and 0.5% monoethanolamine aqueous solution was added and shaken to seal unreacted epoxy groups and dextran. A cellulose gel with immobilized sodium sulfate was obtained. The amount of anionic functional groups introduced by the immobilized dextran sulfate was lOμ-01 per ml of adsorbent.

Example 4 Add 0.0% glycine to 5 ml of the epoxidized gel obtained in Production Example 1.
A solution of 221- dissolved in 1 Gml of water and adjusted to pH 9.8 was added, and the mixture was shaken at room temperature for 24 hours. Thereafter, the gel was filtered, and a 0.5% aqueous monoethanolamine solution was added and shaken to seal unreacted epoxy groups to obtain a cellulose gel with immobilized glycine. The amount of anionic functional group introduced by immobilized glycine was 9 μmol per ml of adsorbent.

Example 5 Add 0.3 taurine to 5 ml of the epoxidized gel obtained in Production Example 1.
A solution prepared by dissolving 7&- in 10 ml of water and adjusting the pH to 9.0 was added thereto, and the mixture was shaken at room temperature for 24 hours. Thereafter, the gel was filtered, and a 0.5% monoethanolamine aqueous solution was added and shaken to seal unreacted epoxy groups to obtain a cellulose gel with taurine immobilized thereon. The amount of anionic functional group introduced by immobilized taurine was 5 μmol per ml of adsorbent.

Example 6 CK gel A3.10ml similar to that used in Production Example 1
After washing with water, suction filtration was carried out, and 6 m of dimethyl sulfoxide was added to this.
1, 2N-NaOH2, 6ml, epichlorohydrin 1
．． 5 ml was added and stirred at 40°C for 2 hours. After the reaction, the gel was filtered and washed with water to obtain a cellulose gel into which epoxy groups were introduced.

6 ml of concentrated ammonia water was added to this and reacted at 40°C for 2 hours to obtain an aminated cellulose gel.

In 5 ml of this gel, 0.2 g of sodium polyacrylate with a molecular weight of 190,000 to 500,000 was dissolved in 10 ml of water, and pna
, add the adjusted solution to s, and then add 1-ethyl-3-(
dimethylaminopropyl) carbodiimide 200p
The mixture was added while maintaining the temperature at 4.5 H and was shaken at 4°C for 24 hours. After the reaction, the gel was filtered and washed with water to obtain a cellulose gel into which polyacrylic acid had been introduced. The amount of anionic functional groups introduced by immobilized polyacrylic acid is 1 m of adsorbent.
It was 14 μmol per liter.

Example 7 Porous cellulose gels were CK Gel A22 (trade name, manufactured by Chisso ■, exclusion limit molecular weight for globular proteins 20 million, particle size 45-1O5I!R) and Cellulofine GCL-200.
0m (product name, manufactured by Chisso ■, exclusion limit molecular weight of globular protein 3 million, particle size 44-105 fi), Cellulofine GCL-1000m (product name, manufactured by Chisso ■,
Exclusion limit molecular weight of globular protein: 600,000, particle size: 44-105
Cellulofine CC (OOs+ (trade name, manufactured by Chisso ■, exclusion limit molecular weight of globular protein 400,000, particle size 4
4 to 105), Cellulofine GC-200 layer (trade name, manufactured by Chisso ■, exclusion limit molecular weight for globular proteins 120,000,
Particle size 45-105 m+), Cellulofine GCL-
Cellulose on which dextran sodium sulfate was fixed was prepared in the same manner as in Production Example 1 and Example 3, except that 90 (trade name, manufactured by Chisso ■, exclusion limit molecular weight of globular protein 35,000, particle size 45 to 105 m+) was used. I got the gel. The amounts of anionic functional groups introduced by immobilized dextran sulfate were 16, 18, and 30.2 per ml of adsorbent, respectively.
It was 4.30.37μ■o1.

Example 8 Example except that an epoxidized cross-linked agarose gel, Epoxy Activated Sepharose CL-6B (trade name, manufactured by Pharmacia Fine Chemicals, exclusion limit molecular weight for globular proteins 4 million, particle size 45-185 m+) was used. Dextran sodium sulfate was immobilized in the same manner as in 3. The amount of anionic functional groups introduced by immobilized dextran sulfate was 20 per ml of adsorbent.
It was μho o1.

Example 9 PP-HG (trade name, manufactured by Mitsubishi Kasei ■, exclusion limit molecular weight for globular proteins: 4 million, particle size: 12
A gel on which dextran sodium sulfate was immobilized was obtained in the same manner as in Production Example 1 and Example 3, except that 0m+) was used. The amount of anionic functional groups introduced by the immobilized dextran sulfate was 9 μs+ol per ml of adsorption.

Example 10 2 g of aminated cellulose gel obtained in the same manner as Example 6
4 g of dextran sodium sulfate having a molecular weight of 500 and a sulfur content of 15% as used in Example 3 was added to 0.
．． A solution dissolved in 8 ml of 1M phosphate buffer (pH 8,0) was added, and the mixture was shaken at room temperature for 18 hours. NaCNB after reaction
After adding 1 g of Hs 2 GI and stirring at room temperature for 30 minutes, the mixture was heated at 40° C. for 4 hours, and the gel was filtered and washed with water to obtain a cellulose gel on which sodium dextran sulfate was fixed. The amount of anionic functional groups introduced by immobilized dextran sulfate was 18 μs+ol per ml of adsorbent.

Example 11 The adsorbent obtained in Examples 1 to 6 and 1O and the carrier CK gel A3 used in Production Example 1 for comparison purposes were 15M
) After washing with O., 15M Tris buffer (pH 7, 4), 0.1 ml of each adsorbent was placed in a polypropylene microtube (capacity: 7 ml) and added with O., 15 M Tris buffer (pH 7.4).
H7,4) was added to bring the total volume to 1 ml. To this was added 0.2 ml of serum containing immune complexes, and the mixture was shaken at 25°C for 2 hours. After completion of this adsorption operation, the gel was centrifuged to sediment, and the immune complex concentration in the collected supernatant was measured by enzyme-linked immunosorbent assay (ELISA method). In other words, to determine the immune complex concentration, add a diluted sample to a C1q-coated plate, perform an antigen-antibody reaction, drop a peroxidase-labeled anti-human immunoglobulin antibody, and perform an enzymatic color reaction using 5LT-210 (trade name, Lab. The measurement wavelength was 486 nm. Measured value is heat denatured 1gG
Standardization was performed using a standard curve created by In Table 1,
The compound group having an anionic functional group immobilized on each adsorbent and the immune complex concentration in the supernatant of each adsorbent are shown converted to the concentration in the original serum taking into account dilution.

From Table 1, it can be seen that the adsorbent formed by immobilizing a compound having an anionic functional group on a water-insoluble porous carrier adsorbs the immune complex. It can be seen that the adsorbent to which dextran sulfate is immobilized has particularly excellent adsorption ability for immune complexes.

[Margin below] Table 1 Example 12 The adsorbents obtained in Examples 3 and 7 to 9 were used, and the amount of serum added was 0.1 ml, but in the same manner as in Example 11, the supernatant was The immune complex concentration was calculated by converting it to the concentration in the original serum taking into account dilution. The results obtained are shown in Table 2 along with the various water-insoluble porous carrier groups used. For comparison, the immune complex concentration of the original serum was measured and found to be 32.2 μg/ml.

From Table 2, Cellulofine CC-700 of Example 7 is a water-insoluble porous carrier with an exclusion limit molecular weight of 400,000 or less.
It can be seen that the immune complex adsorption ability of Cellulose Fine GC-200rn and Cellulose GCL-90 is reduced. On the other hand, the results for cellulose CK gel A3 of Example 3 show that even if the exclusion limit molecular weight is set too high as 50 million, the immune complex adsorption ability tends to decrease.

[Margins below] Table 2 [Effects of the Invention] The adsorbent of the present invention and the removal device using the same have the effect of selectively removing immune complexes from body fluids.

[Brief explanation of the drawing]

FIG. 1 is a graph showing the results of examining the relationship between flow velocity and pressure drop using three types of gels, and FIG. 2 is an overview of an embodiment of the immune complex removal device of the present invention - new aspects. It is a diagram. (Main symbols in the drawing) (1) Two inlets (2) Two outlets (3): Adsorbent (4), (5): Filter (7): Container pressure drop ΔP (k8/am2)

Claims (1)

[Scope of Claims] 1. An adsorbent for an immune complex comprising a compound having an anionic functional group immobilized on a water-insoluble porous carrier. 2. The immune complex adsorbent according to claim 1, wherein the water-insoluble porous carrier has an exclusion limit molecular weight of globular proteins of 400,000 to 60,000,000. 3. The immune complex adsorbent according to claim 1, wherein the water-insoluble porous carrier comprises a compound having a hydroxyl group. 4. The immune complex adsorbent according to claim 1, wherein the compound having an anionic functional group is a polyanionic compound having a plurality of anionic functional groups in one molecule. 5. A container having an inlet and an outlet for a fluid, a filter through which the fluid and the components contained in the fluid can pass through, but not through which the immune complex adsorbent according to claim 1 can pass, and the filter filled in the container. An immune complex removal device consisting of an immune complex adsorbent.