JPH01119264A - Adsorbent and removing device therewith - Google Patents

Abstract

PURPOSE:To selectively adsorb antilipid antibodies substantially without losing effective components in body fluids, by fixing a compound with an anionic functional group to a water-insoluble porous body. CONSTITUTION:An adsorbent of antilipid antibodies is prepared by fixing a compound with an anionic functional group to a water-insoluble porous body. The anionic functional group may be any one or more of the following: sulfate group, sulfonate group, thiol group and phosphate group. The water-insoluble porous body is a compound with a hydroxyl group.

Description

[Detailed description of the invention] [Industrial application field].

The present invention relates to an anti-lipid antibody adsorbent for adsorbing and recovering anti-lipid antibodies from body fluids, and an anti-lipid antibody removing device using the adsorbent.

[Problems to be solved by conventional techniques and inventions] Autoimmune diseases, as the name suggests, are diseases in which antibodies against the constituent components of one's own tissues (hereinafter referred to as autoantibodies) appear, but systemic lupus erythematosus ( In SLE (hereinafter referred to as SLE) and related autoimmune diseases, thrombosis,
In cases such as thrombocytopenia and intrauterine fetal death, antibodies against phospholipid components of cell membranes (hereinafter referred to as antilipid antibodies) appear in body fluids and are thought to be closely related to the pathological condition. The mechanism by which autoantibodies produced are involved in the onset of disease is not necessarily clear, but there is a mechanism by which autoantibodies themselves damage cells, or autoantibodies combine with antigens to form immune complexes that are deposited in tissues. Mechanisms that cause tissue damage have been proposed.

In this case, the generated anti-lipid antibodies bind to phospholipids on platelet membranes, activating or inhibiting their function, inducing thrombosis or thrombocytopenia, respectively, and the endothelium of the placental duct. A mechanism has been proposed that involves binding to phospholipids on cell membranes, inhibiting blood circulation, and inducing fetal death.

Since various symptoms are caused by the anti-lipid antibodies produced in this way or the immune complexes between the antibodies and phospholipids on the cell membrane, controlling the anti-lipid antibodies is extremely important in the treatment of these cases. is important.

Steroids, immunosuppressants, immunomodulators, anti-inflammatory agents, and the like have been widely used in the treatment of SLE for the purpose of suppressing the production of antilipid antibodies.

Among these, steroid drugs are most commonly used, and short-term, high-dose steroid therapy called pulse therapy is often performed. However, since steroids tend to cause side effects even when administered in small doses, it is obvious that short-term, high-dose steroid therapy tends to cause even greater side effects. In addition, these drugs are often used for long periods of time, in which case side effects are more likely to occur, and the dosage must be gradually increased due to drug resistance. There are many cases where it is impossible to use or does not have sufficient effect. In particular, although this is the time when anti-lipid antibodies need to be suppressed most, it is often not possible to use strong therapy using drugs such as pulse therapy or immunosuppressants for the reasons mentioned above.

On the other hand, as an approach different from these drug treatments, attempts have been made to directly remove anti-lipid antibodies from body fluids by extracirculation. The simplest method is so-called plasma exchange therapy, in which the patient's plasma containing anti-lipid antibodies is exchanged with the plasma of a healthy person. This method has shown that anti-lipid antibodies in the blood are significantly reduced and symptoms are improved. However, this method not only requires a large amount of healthy plasma and is expensive, but also involves the risk of infection such as serum hepatitis during the therapeutic treatment, so it has not become widely used.

In plasma exchange therapy, all components in plasma are removed and replaced with healthy plasma, but in order to selectively remove anti-lipid antibodies, 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 high and low molecular weight fractions using a membrane, the high molecular weight fraction containing pathogenic substances is discarded, and the low molecular weight fraction containing albumin, the main protein, is discarded. When returned to the patient, the anti-lipid antibody has a molecular weight of approximately 16
The main component is IgG (immunoglobulin G), which has a similar molecular weight to albumin (molecular weight approximately 60,000), so separation between the two is poor, and a large amount of albumin is also removed when anti-lipid antibodies are removed. It has the disadvantage that all proteins with a molecular weight equal to or higher than that are removed.

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

In view of this situation, the present inventors have conducted intensive research and have discovered an adsorbent that can selectively adsorb only anti-lipid antibodies without losing most of the active ingredients in body fluids, and have completed the present invention. It's arrived.

[Means for Solving the Problems] That is, the present invention provides an anti-lipid antibody adsorbent in which a compound having an anionic functional group is immobilized on a water-insoluble porous material, and a container having an inlet and an outlet for fluid. , a filter that allows passage of a fluid and components contained in the fluid, but not an adsorbent of an anti-lipid antibody, which is formed by immobilizing a compound having an anionic functional group on a water-insoluble porous body;
and an anti-lipid antibody removal device comprising the anti-lipid antibody 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.

The water-insoluble porous body used in the present invention preferably has large diameter continuous pores. In other words, anti-lipid antibodies consist of immunoglobulins such as IgG SI gM (immunoglobulins) and are large molecules with a molecular weight of 180,000 to 900,000. It is necessary to invade the 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. What is the exclusion limit molecular weight?
In gel permeation chromatography, it refers to the molecular weight of the smallest molecular weight of molecules that cannot enter (excluded) the pores.

It is known that the exclusion limit molecular weight varies depending on the target compound, and in general, it has been well investigated for globular proteins, dextran, polyethylene glycol, etc.
It is appropriate to use values obtained using globular proteins (including viruses) that are considered to be most similar to anti-lipid antibodies.

As a result of studies using various water-insoluble porous materials with different exclusion limits, we found that, contrary to expectations, even those with an exclusion limit molecular weight of about 10,000 liters, which is smaller than the molecular weight of anti-lipid antibodies, had a certain degree of adsorption ability. It has become clear that the ability is not particularly great, but rather that the ability is reduced and that proteins other than anti-lipid antibodies are adsorbed, that is, there is an optimal pore size range. In other words, if a water-insoluble porous material with an exclusion limit molecular weight of less than 100,000 is used, the amount of anti-lipid antibody adsorbed is too small to be practical; The use of water-insoluble porous materials similar to the above has been used as a practical adsorbent to some extent. On the other hand, as the exclusion limit molecular weight increases, the adsorption amount of anti-lipid antibodies increases, but eventually reaches a plateau, and when the exclusion limit molecular weight exceeds 80 million, the surface area is too small and the adsorption amount not only decreases noticeably, but also Adsorption of components other than anti-lipid antibodies, that is, non-specific adsorption, increases and selectivity decreases significantly.

Therefore, the exclusion limit molecular weight of the water-insoluble porous material used in the present invention is preferably 10,000 or more and eooo, 0,000 or less, and more preferably 60,000 or more from the viewpoint of a larger selective adsorption capacity.
It is preferable that the number is from 10,000 to 30,000,000.

Next, regarding the porous structure of the water-insoluble porous material, total porosity is preferable to surface porosity, and the pore volume is preferably 20% or more from the viewpoint of a large adsorption capacity. The shape of the water-insoluble porous material is granular, spherical, fibrous, film-like,
Any shape such as hollow fiber can be selected.

When using a particulate water-insoluble porous material, if the particle size is less than 1 liter, the pressure loss will be large;
If the capacity exceeds 500, the capacity of the tracheal tube is small.
It is preferable that it is 0 Is or less.

The water-insoluble porous body 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 anti-lipid antibody. It is preferable that the water-insoluble porous body is hydrophilic rather than hydrophobic, since non-specific adsorption is less likely to occur if it is hydrophilic, and a water-insoluble porous body made of a compound having a hydroxyl group in the molecule is more preferable.

Representative examples of the water-insoluble porous material 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, and styrene-divinylbenzene copolymer; and/or cellulose. Examples include, but are not limited to, porous polymer hard gels made from natural polymers such as .

When the adsorbent of the present invention is used for extracirculation treatment, blood,
Because it is necessary to flow an anti-viscous fluid such as plasma at high speed,
Preferably, a rigid water-insoluble porous body having sufficient mechanical strength not to cause compaction is used. In other words, a hard porous body is one in which a water-insoluble porous body is uniformly packed into a cylindrical column and the relationship between pressure loss and flow rate when an aqueous fluid is passed through is at least 0.3 kg, as shown in the reference example below.
It refers to things that are directly related to /cj.

Any anionic functional group used in the present invention may be used as long as it is negatively charged at around neutral pH. Typical examples of these include carboxyl groups, sulfonic acid groups, sulfone groups, sulfate ester groups, thiol groups,
Examples include, but are not limited to, silanol groups, phosphate ester groups, and phenolic hydroxyl groups.

Among these, carboxyl groups, sulfonic acid groups, sulfate ester groups, thiol groups, and phosphate ester groups are preferred because they have a strong affinity for anti-lipid antibodies.

As a compound having an anionic functional group, 1
It may be a monoanionic compound having one anionic functional group or a polyanionic compound having multiple anionic functional groups. Polyanionic compounds are preferred because they have a high affinity for anti-lipid antibodies and are easy to introduce many anionic functional groups into a unit amount of porous material. Among these, polyanionic compounds having a molecular weight of too much or more are preferred in terms of affinity and the amount of anionic functional group introduced. The polyanion compound may have one or two types of anionic functional groups.

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 a water-insoluble porous material in which a compound having an anionic functional group is fixed. There are various methods of introducing an anionic functional group into an adsorbent to achieve a fixed state of a compound having such an anionic functional group. The methods include (1) forming an adsorbent by polymerization using an anionic functional group or a compound containing a functional group that can be easily converted into an anionic functional group as a monomer or a crosslinking agent; (2) forming an adsorbent using an anionic functional group; (3) A method of fixing an anionic functional group to a water-insoluble porous body by directly reacting the compound that forms the anionic functional group with the water-insoluble porous body. Examples include a method of immobilizing a compound having such a compound.

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 of fixing a compound containing an anionic functional group to a water-insoluble porous material, includes methods of physical adsorption, methods of ionic bonding, and methods of fixing by covalent bonding. However, in order to use adsorbents for therapeutic purposes, it is important that the anionic functional group-containing compound does not detach during sterilization or treatment, so covalent bonding methods that allow for strong immobilization are recommended. 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 compounds containing anionic functional groups having these functional groups, such as sulfanilic acid, 2-aminoethyl hydrogen sulfate, terephthalic acid, phosphorylethanolamine, glucose 6-phosphoric acid, and ethanedithiol described in the examples. is one example.

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, among which polyhydric alcohols The sulfate-esterified product, especially the sulfate-esterified product of sugars, contains both the sulfate ester group and the functional group necessary for immobilization, and has high biocompatibility and activity, and furthermore, sulfated polysaccharides are easily hydrated. It is particularly preferred because it can be fixed to an insoluble porous body.

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

The amount of anionic functional group introduced is preferably 0.01 u mol or more and 10 mmol or less per ml of adsorbent. 0. If the amount of O is less than 1 μmol, the adsorption capacity is insufficient, and if it exceeds low mol, there will be too much non-specific adsorption, making it difficult to put it to practical use.

More preferably, the amount of anionic functional group introduced is 1 μsol or more and 100 μsol or less.

There are various methods for removing anti-lipid antibodies from body fluids using the adsorbent of the present invention, and any method may be used. can pass through the filter, but cannot pass through the anti-lipid antibody adsorbent, which is a water-insoluble porous material with a compound having an anionic functional group fixed thereto, and from the anti-lipid antibody adsorbent filled in the container. The method of passing body fluid through the anti-lipid antibody removing device is simple and preferable. In Fig. 2, (1) and (2) are the inlet and outlet of each fluid, (3) 1 ± the adsorbent of the present invention, (4) and (5) are the fluid and the components contained in the fluid. A filter or mesh through which the adsorbent of the present invention can pass, (6) a column, and (7) a container. Here, the filter (4) 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 Biogol A5m (trade name, manufactured by Bio-Rad, Inc.,
(particle size: 50-100 mesh), synthetic polymer gel, Toyovar HW85 (product name, manufactured by Toyo Soda Kogyo ■, particle size: 50-10100u), and porous cellulose gel, Cellulofine CC-700 (product name, manufactured by Chisso ■) , particle size 45 to 100 ρ) was uniformly packed, water was passed through the column using a veristatic pump, and the relationship between the flow rate and the pressure drop ΔP was determined.The results are shown in Figure 1. As is clear, agarose gel, which is a soft gel, undergoes compaction when the flow rate exceeds a certain level, and the flow rate does not increase even if the pressure is increased, whereas hard gels such as Toyopearl and Cellulofine are almost proportional to the increase in pressure. The flow rate increases.

Production Example 1 CK Gel A3 (trade name,
Made by Chisso ■, exclusion limit molecular weight of globular protein 50 million,
Particle size: 83 to 125) To 100 ml were added 180 ml of water B and 2M Na0, and the mixture was stirred at 45°C for 1 hour.

After stirring, add 25ml of shrimp chlorohydrin and
After the reaction was completed by stirring at 5° C. for 2 hours, the gel was filtered and washed with water to obtain an epoxidized cellulose gel (hereinafter referred to as epoxidized gel).

Example 1 A solution of 0.14 g of sulfanilic acid dissolved in 7.5 ml of water was added to the epoxidized gel loml obtained in Production Example 1, and after adjusting the pH of the solution to 10 by adding 2M Na011, the solution was heated at 45°C. It was left for 20 hours. After the reaction is complete,
The gel was filtered and washed with water to obtain a cellulose gel on which sulfanilic acid was fixed. The amount of anionic functional groups introduced by immobilized sulfanilic acid is 1O per ml of adsorbent.
It was μff1O1.

Example 2 2-aminoethyl hydrogen sulfate instead of sulfanilic acid
．． A cellulose gel on which 2-aminoethyl hydrogen sulfate was immobilized was obtained in exactly the same manner as in Example 1, except that 11 g was used. The amount of anionic functional group introduced by the immobilized 2-aminoethyl hydrogen sulfate was 10 μmol per ml of adsorbent.

Example 3 A cellulose gel on which phosphorylethanolamine was immobilized was obtained in exactly the same manner as in Example 1, except that phosphorylethanolamine o,ttg was used instead of sulfanilic acid. The amount of anionic functional group introduced by immobilized phosphorylethanolamine is 1 ml of adsorbent.
It was 1Oμl 1O1 per.

Example 4 1,2-ethanedithiol 0 instead of sulfanilic acid
．． A cellulose gel on which 1,2-ethanedithiol was fixed was obtained in exactly the same manner as in Example 1 except that 08 [ was used. The amount of anionic functional group introduced by immobilized 1,2-ethanedithiol was 10 μmol per ml of adsorbent.

Example 5 To 10 ml of the epoxidized gel obtained in Production Example 1,
000, 5g1 of dextran sodium sulfate containing 18% sulfur and 8ml of water were added, and further 2M NaOH
After adjusting the pH of the solution to 10 by adding
I left it for a while. After the reaction, the gel was filtered and washed with water, and 0.5%
A monoethanolamine aqueous solution was added and left at room temperature for 20 hours to seal unreacted epoxy groups. After the reaction was completed, the gel was filtered and washed with water to obtain a cellulose gel on which dextran sodium sulfate was fixed. The amount of anionic functional groups introduced by immobilized dextran sulfate is
It was 29 μmol per liter.

Production Example 2 A solution of 0.5 g of ethylenediamine dissolved in 30 ml of water was added to 40 ml of the epoxidized gel obtained in Production Example 1, and the mixture was left at 45°C for 20 hours. After the reaction was completed, the gel was filtered and washed with water to obtain a cellulose gel into which amino groups had been introduced (hereinafter referred to as aminated gel).

Example 6 To 10 ml of the aminated gel obtained in Production Example 2, 10 ml of water was added, followed by glucose 6-phosphate barium salt Ig, and then 0.5 ml of 2M NaOH was added for 1 hour.
It was kept at 5°C. 40.1 g of NaBII was added to this and left to stand at room temperature for 24 hours. After the reaction was completed, the gel was filtered and washed with water to obtain cellulose on which glucose 6-phosphate was fixed. The amount of anionic functional groups introduced by the immobilized gelcoast phosphoric acid was 10 μmol per ml of adsorbent.

Example 7 10 ml of the aminated gel obtained in Production Example 2 was suspended in N,N-dimethylformamide (DMF) to make a total volume of 18 ml.
And so. After dissolving 0.27 g of terephthalic acid therein, 1 g of a condensation reagent (N,N'-dicyclohexylcarbodiimide) was added and stirred at room temperature for 124 hours. After the reaction was completed, the gel was filtered and washed with DMF, ethanol, and water in this order to obtain a cellulose gel on which terephthalic acid was fixed.

The amount of anionic functional groups introduced by immobilized terephthalic acid was 10 μmol per ml of adsorbent.

Example 8 To 10 ml of the aminated gel obtained in Production Example 2, 5 g of dextran sodium sulfate of the same kind as that used in Example 5 and 8 ml of water were added, and further 5 ml of 2M Na011O was added.
1 was added and kept at 45°C for 1 hour. This has Na1311
40.1 g was added and left at room temperature for 24 hours. After the reaction was completed, the gel was washed point by point with water to obtain a cellulose gel on which dextran sodium sulfate was fixed.

The amount of anionic functional groups introduced by immobilized dextran sulfate was 39 μmol per ml of adsorbent.

Production Example 3 Porous cellulose gel of the same type as that used in Production Example 1 (
40 ml of CK gel A 3 ) was suspended in heptane to make a total volume of 70 ml. 20% N1 Kame Off for this
l Oml and nonionic surfactant Tween 20
(Product name, manufactured by Bio-Rad) 40? Add fi and 40
Shake at ℃ for 30 minutes. followed by epichlorohydrin lo
ml was added and shaken at 40°C for 6 hours. After the reaction was completed, the gel was filtered and washed with ethanol and water in that order to obtain an epoxidized gel.

Example 9 A cellulose gel on which dextran sodium sulfate was fixed was obtained in exactly the same manner as in Example 5, except that the epoxidized gel obtained in Production Example 3 was used instead of the epoxidized gel obtained in Production Example 1. The amount of anionic functional groups introduced by immobilized dextran sulfate is
It was 38 μmol per ml.

Example 1O Polyacrylic acid having an amino group at one end (molecular weight 1000) was added to 10 ml of the epoxidized gel obtained in Production Example 3.
Add Ig dissolved in 5 ml of water, and add 2M
1 ml of Na0II was added and left at room temperature for 48 hours. After the reaction was completed, the gel was filtered and washed with water to obtain a cellulose gel on which polyacrylic acid was fixed. The anionic functional group introduced by immobilized polyacrylic acid is 1 m
It was 560 μmol per liter. Polyacrylic acid having an amino group at one end can be produced by a low polymerization reaction of acrylic acid using 2-aminoethanethiol as a chain transfer agent and α,α″-azobisisobutyronitrile (AIBN) as an initiator. (Journal of the Chemical Society of Japan, 19
77, No. 1, +l [see pages 1-92, "Synthesis of 2-hydroxyethyl-methacrylate-styrene type AIIA block copolymer and its structure and wetting", Mitsuo Okano et al.).

Example 11 Polyacrylic acid having an amino group at one end (molecular weight 1
Example 10 except that polyacrylic acid (molecular weight approximately 10,000) tg having an amino group at one end was used instead of 000).
Cellulose gel with polyacrylic acid immobilized was obtained in exactly the same manner as above. The anionic functional groups introduced by the immobilized polyacrylic acid were 5 Bm mol/ml adsorbent. Polyacrylic acid having an amino group at one end was synthesized according to a method similar to that described in Example 10.

Production Example 4 An aminated gel was obtained in exactly the same manner as Production Example 2, except that 0.12 g of ethylenediamine dissolved in 5 ml of water was added to 10 ml of the epoxidized gel obtained in Production Example 3.

Example 12 A cellulose gel on which dextran sodium sulfate was fixed was obtained in exactly the same manner as in Example 8, except that 10 ml of the aminated gel obtained in Production Example 4 was used instead of the aminated gel obtained in Production Example 2. The amount of anionic functional groups introduced by immobilized dextran sulfate is
It was 76μs+ol per.

Production Example 5 7, Porous cellulose gel CK gel A22 (trade name, product name, manufactured by Chisso ■, globular protein exclusion limit molecule m 3
0 million, particle size 53-125 uta) 25 ml
Add 10 ml of water and 130 ml of 2M Na0 to the
Shake at 40°C for 20 minutes.

To this was added 12 ml of epichlorohydrin, and the mixture was shaken at 40°C for 3 hours. After the reaction was completed, the gel was filtered and washed with water to obtain an epoxidized gel.

Production Example 6 CK gel A32 (trade name, porous cellulose gel)
Manufactured by Chisso ■, exclusion limit molecular weight of globular protein 20 million,
Particle size 53-125. um) 25 ml, 15 ml of water, 121 ml of 2M Na0 and 7 ml of shrimp chlorohydrin.
．． An epoxidized gel was obtained in the same manner as in Production Example 5 except that 1 ml was used.

Production Example 7 Cellulofine GCL-2, a porous cellulose gel
25
An epoxidized gel was obtained in the same manner as in Production Example 5, except that 25 ml of water, 15 ml of 1.2M NaO]l, and 5 ml of shrimp chlorodrine were used.

Production Example 8 Cellulofine GCL-1, a porous cellulose gel
00h (Product name, manufactured by Chisso ■, exclusion limit molecular weight of globular protein GO 10,000, particle size 44~lQ5AIm) 25m
1.

An epoxidized gel was obtained in the same manner as in Production Example 5, except that 25 ml of water, 11.75 ml of 2M Na01111.75 ml and 4 ml of shrimp chlorodrine were used.

Production Example 9 Cellulofine CC-70, a porous cellulose gel
0m (Product name, manufactured by Chisso ■, globular protein exclusion limit molecule FF 1.4 million, particle size 44-105 times) 25m1
.

25ml of water, 2M NaO! An epoxidized gel was obtained in the same manner as in Production Example 5, except that 8.75 ml of shrimp chlorohydrin and 3 ml of shrimp chlorohydrin were used.

Production Example 1O Cellulofine CC-20, a porous cellulose gel
0m (Product name, manufactured by Chisso ■, exclusion limit molecular weight of globular protein 120,000, particle size 44-105m+) 25m1
.

An epoxidized gel was obtained in the same manner as in Production Example 5, except that 25 ml of water, 7 ml of 1.2 M NaOH, and 2.5 ml of shrimp chlorohydrin were used.

Example 13 Epoxidized gel obtained in Production Example 5 (gel A22 in C) 20
ml", 10 g of dextran sodium sulfate of the same kind as that used in Example 5 and water were added to bring the total volume to 33 ml. 2M Na011 was added to bring the I'11 of the solution to 10
．． After adjusting the temperature to 0, the temperature was left at 45° C. for 17 hours. After the reaction was completed, the gel was filtered and washed with water, and a 0.5% monoethanolamine aqueous solution was added thereto and left at room temperature for 20 hours to seal unreacted epoxy groups. After the reaction was completed, the gel was filtered and washed with water to obtain a cellulose gel on which dextran sodium sulfate was fixed. The amount of anionic functional groups introduced by immobilized dextran sulfate was 31 μmol per ml of adsorbent.

Example 14 Epoxidized gel obtained in Production Example 6 (CK Gel Hachihe 2) 20
Example 13 was carried out in the same manner as in Example 13, except that lQg of dextran sodium sulfate of the same type as that used in Example 5 and water were added to make the total volume 3θml, and the pl+ of the solution was adjusted to 9.9. , a cellulose gel with immobilized sodium dextran sulfate was obtained. The amount of anionic functional groups introduced by immobilized dextran sulfate was 27 μa+ol per ml of adsorbent.

Example 15 Epoxidized gel obtained in Production Example 7 (GCL-200011
) to 20 ml, add 9.3 g of dextran sodium sulfate of the same type as that used in Example 5 and water, and bring the total volume up to 3.
A cellulose gel on which dextran sodium sulfate was immobilized was obtained in the same manner as in Example 13, except that the volume was 5 ml and the pl+ of the solution was adjusted to 9.3. The amount of anionic functional groups introduced by immobilized dextran sulfate was 30 μmol per ml of adsorbent.

Example 1B Epoxidized gel obtained in Production Example 8 (GCL-1000m)
To 20 ml, add 9.3 g of dextran sodium sulfate of the same type as that used in Example 5 and water to make a total volume of 35 ml.
A cellulose gel on which dextran sodium sulfate was immobilized was obtained in the same manner as in Example 13, except that the pH of the solution was adjusted to 1 and the pH of the solution was adjusted to 9.3. The amount of anionic functional group introduced by the immobilized dextran sulfate was 30 μmol per ml of shield absorber.

Example 17 Epoxidized gel obtained in Production Example 9 (GC-700111)
To 20 ml, add 9.0 g of dextran sodium sulfate of the same type as that used in Example 5 and water to make a total volume of 36 ml.
A cellulose gel on which dextran sodium sulfate was immobilized was obtained in the same manner as in Example 13, except that the pH of the solution was adjusted to 1 and the pH of the solution was adjusted to 9.3. The amount of anionic functional groups introduced by immobilized dextran sulfate was 30 μmol per ml of adsorbent.

Example 18 Epoxidized gel (GO-200m) 2 obtained in Production Example 1O
0 ml, add 9.0 g of dextran sodium sulfate of the same type as that used in Example 5 and water to make a total volume of 30 ml.
De1: Cellulose gel on which sodium chytran sulfate was fixed was obtained in the same manner as in Example 13, except that the pH of the solution was adjusted to 9.2. The amount of anionic functional groups introduced by immobilized dextran sulfate was 32 μs per ml of adsorbent.
It was +ol.

Example 19 After washing the adsorbents obtained in Examples 1 to 12 with 0.5M phosphate buffer (pH 7,4), 0.1 ml of each adsorbent was washed.
each in a polypropylene microtube (capacity 1.5
ml), and 0.5M phosphate buffer (pH 7.4) was added to bring the total volume to 1 ml. 0 and 2 ml of serum containing anti-lipid antibodies were added thereto, and the mixture was shaken at 37°C for 2 hours. After this adsorption operation is completed, the gel is precipitated by centrifugation, and the anti-lipid antibody titer in the collected supernatant is determined by enzyme immunoassay (E
It was measured by LISA method). The anti-lipid antibody titer was determined by adding the diluted sample to a plate coated with calciolipin, performing an antigen-antibody reaction, adding peroxidase-labeled anti-human immunoglobulin antibody, and performing an enzyme coloring reaction with C5-93.
0 (trade name, manufactured by Shimadzu Corporation). Table 1 lists the names of compounds with anionic functional groups immobilized on each adsorbent, and the anti-lipid antibody titer in the supernatant after the adsorption operation with respect to the anti-lipid antibody titer in the original serum of each adsorbent. Expressed as relative antibody titer in percentage.

Table 1 shows that the adsorbent to which dextran sulfate or polyacrylic acid is immobilized has particularly excellent anti-lipid antibody adsorption ability.

Example 20 Using the adsorbents obtained in Examples 5 and 13 to 18, relative antibody titers were determined in the same manner as in Example 19, except that the amount of serum added was 0.15 ml. The results obtained are shown in Table 2 along with various water-insoluble porous materials used.

Table 2 shows that the anti-lipid antibody adsorption ability of adsorbents using Cellulofine GC-700m and Cellulofine GC-200a+, which are porous materials with exclusion limit molecular weights of 400,000 or less, is slightly lower. Conversely, it can be seen that even if the exclusion limit molecular weight is set too high as 50 million, the anti-lipid antibody adsorption ability tends to decline.

Table 1 [Effects of the Invention] The adsorbent of the present invention and the removal device using the same have the effect of selectively removing anti-lipid antibodies from body fluids.

[Brief explanation of the drawing]

Figure 1 is a graph showing the results of examining the relationship between flow rate and pressure drop using three types of gels, and Figure 2 is a schematic cross-sectional view of one embodiment of the anti-lipid antibody removal device of the present invention. be. (Main symbols in the drawing) (1) Two inlets (2): Outlets (3): Adsorbents (4), (5): Filters (7): Container 21?1 Figure Pressure loss ΔP (Kq/cm” )

Claims (1)

[Scope of Claims] 1. An anti-lipid antibody adsorbent comprising a water-insoluble porous material immobilized with a compound having an anionic functional group. 2. The antilipid according to claim 1, wherein the anionic functional group is at least one selected from the group consisting of a sulfuric acid ester group, a sulfonic acid group, a carboxyl group, a thiol group, and a phosphoric acid ester group. Antibody adsorbent. 3. The anti-lipid antibody 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. 4. The anti-lipid antibody adsorbent according to claim 1, wherein the water-insoluble porous body is made of a compound having a hydroxyl group. 5. A container having an inlet and an outlet for a fluid, through which the fluid and the components contained in the fluid can pass, but an anti-lipid antibody adsorbent consisting of a water-insoluble porous body with a compound having an anionic functional group immobilized thereon is An anti-lipid antibody removal device comprising an impermeable filter and an adsorbent for the anti-lipid antibody filled in the container.