JPH11396A - Device for removing anti-lipid antibody - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an anti-lipid antibody removal device for suctionally removing or suctionally recovering only anti-lipid antibodies in a selective manner with little loss of effective components in humor. SOLUTION: A device for removing anti-lipid antibodies is used which comprises a container 7 having a fluid inlet 1 and outlet 2, a filter passing fluid and components contained in the fluids but not passing an anti-lipid antibody adsorbent 3 obtained when a polyanionic compound having an anionic functional group, with a molecular weight of 1000 or more, except DNA, is fixed to a water-insoluble porous body, and the anti-lipid antibody adsorbent 3 packed in the container 7.

Description

DETAILED DESCRIPTION OF THE INVENTION [0001] BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an anti-lipid antibody
Of anti-lipid antibody for adsorption removal or adsorption recovery
Related to the leaving device. [0002] 2. Prior Art and Problems to be Solved by the Invention
An autoimmune disease is, as its name implies, the composition of its own tissue.
For which antibodies to the minute (hereinafter referred to as autoantibodies) appear
The patient has systemic lupus erythematosus (hereinafter referred to as SLE)
And related autoimmune diseases, thrombosis,
For cases such as thrombocytopenia and intrauterine fetal death
Is an antibody against the phospholipid component of the cell membrane (hereinafter referred to as anti-lipid
Antibodies) appear in body fluids and are closely related to their pathology
It is believed that there is. Autoantibodies produced are sick
Although the mechanism involved in the development is not always clear, autoantibodies
The mechanism that damages cells or autoantibodies
By binding to form immune complexes and depositing in tissues
Mechanisms that cause organizational damage have been proposed. The case
In this case, the generated anti-lipid antibody is
Binds to lipids, activates or inhibits their function,
Inducing thrombosis or thrombocytopenia, respectively.
In addition, it binds to phospholipids on the endothelial cell membrane of
Mechanisms that inhibit fetal death and induce fetal death
You. [0003] The anti-lipid antibody thus produced or the anti-lipid antibody
Immune complex between antibody and phospholipid on cell membrane
Can cause severe symptoms,
The control of anti-lipid antibody is very important for treatment. The purpose of suppressing the production of anti-lipid antibodies conventionally
Steroids, immunosuppressants, immunomodulators, anti-inflammatory agents
Are widely used for the treatment of SLE. Above all,
Steroids are the most commonly used, pulsed therapy
Short-term, high-dose steroid therapy called
Is being done. However, steroids can be used in small doses.
Because it is easy to cause side effects even if given, steroids
More severe side effects with short-term ultra-high dose therapy
It is self-evident that it will be easier for you. In addition, these drugs
Is often used for a long time,
In the meantime, side effects are more likely to occur and drug resistance
Because the dose often needs to be increased gradually,
In some cases these drugs are not available,
There are many cases where the effect is not sufficient. In particular, the case
Appears when anti-lipid antibody suppression is most needed
Pulse therapy for the above reasons
Powerful therapies using drugs such as drugs and immunosuppressants
If not, there are many cases. [0005] On the other hand, an approach other than these drug therapies is used.
Anti-lipid antibodies in body fluids by direct external circulation
Attempts have been made to leave. The easiest one
The method involves replacing a patient's plasma containing anti-lipid antibodies with plasma from a healthy individual.
This is a so-called plasma exchange therapy. This method
Anti-lipid antibodies in the blood dropped significantly and symptoms improved.
Have been. However, this method requires a large amount of healthy plasma
Is not only expensive and expensive, but also during the treatment
Widely spread due to the risk of infection such as serum hepatitis
Has not been reached. In plasma exchange therapy, all components in the plasma
It is removed and replaced with healthy plasma.
On the other hand, it selectively removes anti-lipid antibodies, which are pathogens
Plasma separation for the purpose of separating pathogens by molecular size
The membrane method was developed. In this method, plasma is polymerized by a membrane
High molecular weight fraction and low molecular weight fraction
High molecular weight fraction is discarded and albumin, the main protein,
Return the low molecular weight fraction contained to the patient
Is mainly IgG (immunoglobulin G) with a molecular weight of about 160,000
Yes, because the molecular weight is close to albumin (molecular weight about 60,000)
The separation between the two is poor, and when removing anti-lipid antibodies
Bumin is also removed in large amounts, and at the same
There is a disadvantage that all proteins of a high molecular weight are removed. [0007] Therefore, anti-lipid antibodies,
More selective removal, almost no other useful components in body fluids
There has been a desire for a means of removal that will not be lost. [0008] [Means for Solving the Problems] The present inventors have studied such a technique.
As a result of intensive studies in consideration of the
Selectively adsorbs only anti-lipid antibodies without losing much
The present inventors have found an adsorbent that has completed the present invention. That is, the present invention provides a fluid inlet and a fluid inlet.
Container having an outlet, fluid and components contained in the fluid
Can pass, but the water-insoluble porous material has an anionic functional group.
A polyanion compound having a molecular weight of 1,000 or more,
Adsorption of anti-lipid antibody immobilized but excluding DNA
Filters that cannot pass through the body, and filled in the container
Of an anti-lipid antibody comprising an adsorbent of the anti-lipid antibody
Related to the leaving device. [0010] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS In the present specification, body fluid refers to blood, plasma, and blood.
Qing, ascites, lymph, intra-articular fluid and obtained from these
Fractions and other biologically derived liquid components.
U. The water-insoluble porous material used in the present invention has a large size.
Those having continuous pores of various diameters are preferred. Sand
Anti-lipid antibodies are IgG, IgM (immunoglobulin)
M) and immunoglobulins having a molecular weight of 16 to 9
Because it is a macromolecule of 100,000, it can be efficiently adsorbed
That the anti-lipid antibody can easily enter the porous body
is necessary. There are various methods for measuring the pore size.
The method is most often used, but the hydrophilic porous
It is difficult to apply in some cases. Alternative pore diameter guide
The exclusion limit molecular weight is often used, hydrophilic porous body,
It can be applied to any hydrophobic porous body. Exclusion limit molecular weight
Is a book (for example, Hiroyuki Hatano, Toshihiko Hanai, experimental high-speed
Liquid chromatography, chemical doujin)
Like in gel permeation chromatography
Smallest molecule that cannot enter (exclude)
The molecular weight of a substance having a high molecular weight. The exclusion limit molecular weight depends on the target compound.
Are known to be different, generally globular proteins, dex
Investigate well for tolan, polyethylene glycol
And seems most similar to anti-lipid antibodies
Values obtained using globular proteins (including viruses)
It is appropriate to use Various water-insoluble porous materials having different exclusion limits
As a result, the exclusion limit molecular weight was unexpectedly higher than expected.
It is about 100,000 smaller than the molecular weight of lipid antibody
Level of adsorption capacity, and the larger the pore size, the higher the capacity.
Not big, but rather reduced ability or anti-lipid antibody
Other proteins are adsorbed, that is,
It was revealed that a range existed. That is, 10
Using a water-insoluble porous material with an exclusion limit molecular weight of less than 10,000
In the case of tobacco, the adsorption amount of anti-lipid antibody is small and it cannot be used
However, the exclusion limit molecular weight is 100,000 to 150,000 and anti-lipid
Even with water-insoluble porous materials close to the molecular weight of the body
A practically usable adsorbent was obtained. On the other hand, exclusion limit molecules
As the amount increases, the adsorption amount of anti-lipid antibody increases
Eventually it reached a plateau and the exclusion limit molecular weight reached 60 million
Above this, the surface area is too low and the adsorption is noticeably reduced
Not only absorbs components other than the anti-lipid antibody of interest.
Deposition, that is, nonspecific adsorption increases and selectivity is remarkable
Lowers. Therefore, the water-insoluble porous material used in the present invention
Exclusion limit molecular weight of the polymer is 100,000 to 60,000,000
Or less, and more preferably a larger selective adsorption capacity.
600,000 to 600,000, even 600,000
It is good that it is more than 30 million. Next, the porous structure of the water-insoluble porous body will be described.
Therefore, total porosity is preferred over surface porosity, and pore volume is
The product must be at least 20% in terms of large adsorption capacity.
Is preferred. The shape of the water-insoluble porous body is granular, spherical
Any shape such as shape, fiber shape, film shape, hollow fiber shape
You can choose. Using water-insoluble porous particles
If the particle size is less than 1 μm, the pressure drop
Adsorption capacity when loss is large and exceeds 5000μm
Is smaller than 1 μm and smaller than 5000 μm
Is preferred. The water-insoluble porous material used in the present invention is an organic
May be either acidic or inorganic, but the desired anti-lipid
Low adsorption of body fluid components other than antibodies (so-called non-specific adsorption)
None is preferred. Non-specific adsorption is less when hydrophilic
Water-insoluble porous material is more hydrophilic than hydrophobic
Compounds having a hydroxyl group in the molecule.
A water-insoluble porous body made of a material is more preferable. Representative of the water-insoluble porous material used in the present invention
Examples are agarose, dextran, polyacrylic
Soft porous materials such as amide, porous glass, porous silicon
Inorganic porous material such as kagel, polymethylmethacrylate
G, polyvinyl alcohol, styrene-divinylbenze
Synthetic polymers such as copolymers and / or cellulose
Porous polymer hardgel made from natural polymers such as
But not limited to these.
No. The adsorbent used in the present invention is used for extracorporeal circulation treatment
Flow high-viscosity fluids such as blood and plasma at high speed.
Sufficient machine that does not cause consolidation
It is preferable to use a hard water-insoluble porous material having
New The hard porous body is, as shown in Reference Examples below,
Water-insoluble porous material is uniformly packed in a cylindrical column,
When the fluid circulates, the relationship between pressure loss and flow rate is small.
At least 0.3kg / cm^TwoUp to a linear relationship
U. The anionic functional group used in the present invention has a pH of
Any functional group that is negatively charged near neutral
One can also be used. Representative examples of these are
A sil group, a sulfonic group, a sulfone group, a sulfate group,
Thiol group, silanol group, phosphate group, pheno
But not limited to these.
Do not mean. Above all, carboxyl group, sulfonic acid
Group, sulfate group, thiol group and phosphate ester
The groups have strong affinity for anti-lipid antibodies and are preferred. As a compound having an anionic functional group
Is a monoaniline having one anionic functional group in the molecule.
Possessing multiple anionic functional groups rather than on compounds
Lianion compounds are preferred. Polyanion compounds are anti-
High affinity for lipid antibodies and unit volume porosity
It is easy to introduce many anionic functional groups into
New Polyanion with a molecular weight of 1000 or more
Compounds are preferred in terms of affinity and introduction of anionic functional groups.
No. The polyanionic compound has one anionic functional group.
The seeds may be used, or two or more kinds may be used. Representative of the polyanion compound used in the present invention
Examples include polyacrylic acid, polyvinyl sulfate, polyvinyl
Nyl sulfonic acid, polyvinyl phosphoric acid, polystyrene sulf
Honic acid, polystyrene phosphoric acid, polyglutamic acid, poly
Aspartic acid, polymethacrylic acid, polyphosphoric acid, styrene
Synthetic polyanion compound such as len-maleic acid copolymer
And heparin, dextran sulfate, chondroitic
, Chondroitin sulfate, phosphomannan, chitin, chi
Polysaccharides containing anionic functional groups such as tosan
However, it is not limited to these. [0023] The affixed to the adsorbent used in the present invention.
Polyaniline having a nonionic functional group and a molecular weight of 1,000 or more
ON compounds, except for DNA, may be of one type
Alternatively, two or more types may be used. The adsorbent used in the present invention is a water-insoluble porous material.
Having a molecular weight of 1000 or more having an anionic functional group
Ryanion compound, except DNA, immobilized
It refers to the state. Has such an anionic functional group
Polyanion compounds with a molecular weight of 1,000 or more and others
The fixed state of the compound having an anionic functional group
The method of introducing an anionic functional group into an adsorbent to obtain
And can be introduced in any way, but
How to enter (1) Anionic functional group or easily anionic functional
A compound containing a functional group that can be converted to a group is a monomer
To form an adsorbent by polymerization
(2) a method of adding a compound containing an anionic functional group to water
Method for fixing to insoluble porous material, (3) anionic agent
The water-insoluble porous material is directly reacted with the
By adding an anionic functional group to the water-insoluble porous body.
And a method of fixing a compound having a group. The anionic agent used in the method (1)
Functional groups or functional groups that can be easily converted to anionic functional groups
As a representative example of a monomer or a crosslinking agent containing a group
Are acrylic acid and its esters, methacrylic acid and
And its esters, styrene sulfonic acid, etc.
Are not limited to these. The method (2), ie, an anionic functional group
For immobilizing a compound containing water on a water-insoluble porous body
The methods include physical adsorption and ionic bonding.
Method, method of fixing by covalent bond, etc.
Method may be used, but the use of adsorbents for therapeutic purposes
Is a compound containing an anionic functional group during sterilization or during treatment.
It is important that the object does not come off, so it can be firmly fixed.
An efficient covalent bonding method is preferred. A compound containing an anionic functional group by a covalent bond
When fixing the product, the anionic functional group-containing compound
Has functional groups that can be used for immobilization other than anionic functional groups
Preferably. Representative examples of functional groups that can be used for fixing
Is amino group, amide group, carboxyl group, acid anhydride
Group, succinylimide group, hydroxyl group, thiol group, aldehyde
Hyd group, halogen group, epoxy group, silanol group, etc.
But not limited to these. Anionic functional groups having these functional groups
Although there are many contained compounds, sulfanyl described later
Acid, 2-aminoethyl hydrogen sulfate, terephthalic acid, phospho
Lilethanolamine, glucose 6-phosphate, ethane
Dithiol is one example. Further, a compound containing an anionic functional group
Of the compounds containing sulfate groups
Is to make hydroxyl group of alcohol, sugar, glycol etc.
Compound sulfates, among these,
Partially sulfated esters of polyhydric alcohols, especially sugars
Sulfate ester of sulfate group, necessary for fixing
It contains both functional groups and is biocompatible and active
Both are high, and sulfated polysaccharides are easily water-insoluble and porous
It is particularly preferable because it can be fixed to a substrate. Next, the method (3), ie, anionic
The compound forming the functional group reacts with the water-insoluble porous body.
By adding an anionic functional group to the water-insoluble porous body.
Anionic functional group by immobilizing compound with group
As a typical example of the method for performing the treatment, sulfuric acid
There is a reaction for introducing a ter group. In this case, hydroxyl
Group containing water-insoluble porous material, chlorosulfonic acid, concentrated sulfuric acid
Which reagent reacts directly with the sulfate group
Can be introduced. The amount of anionic functional group introduced depends on the amount of adsorption
0.01 μmol or more and 10 mmol or less per 1 ml of body
Below is preferred. Adsorption capacity when less than 0.01μmol
Is not sufficient and exceeds 10 mmol, non-specific absorption
Wearing too much makes it difficult to put it to practical use. More preferred
New anionic functional group introduction amount is 1μmol or more and 100μ
mol or less. [0033] Using the adsorbent used in the present invention, anti-body
There are various methods for removing lipid antibodies.
A container having an inlet and an outlet for fluid may be used.
Vessels, fluids and components contained therein can pass through,
Molecular weight 1 having anionic functional group in water-insoluble porous material
000 or more polyanionic compounds, excluding DNA
Adsorbent of immobilized anti-lipid antibody cannot pass through
Filter, and the anti-fat filled in the container
Body fluid to the anti-lipid antibody removal device consisting of
The method of passing the solution is simple and preferable. FIG. 1 shows an anti-lipid antibody removing apparatus according to the present invention.
1 shows a schematic sectional view of an embodiment. 1 and 2 in FIG.
The inlet and outlet of each fluid, 3 is the adsorbent of the present invention, 4
And 5 can pass through the fluid and the components contained in the fluid
However, the adsorbent used in the present invention cannot pass through a filter or
Is a mesh, 6 is a column, and 7 is a container. Where the fluid
The filter 4 on the inlet side may not be present. Hereinafter, the removing apparatus of the present invention will be described by way of examples.
As will be described in more detail below, the present invention is limited to only these examples.
It is not specified. Reference example Glass cap fitted with a filter with a pore size of 15 μm at both ends
Agarose on ram (inner diameter 9mm, column length 150mm)
Gel Biogel A5m (trade name, manufactured by Bio-Rad, Inc.
Particle size of 50-100 mesh), made of synthetic polymer
Le, Toyopearl HW65 (trade name, Toyo Soda Kogyo Co., Ltd.)
, 50-100 μm particle size), and porous cellulose
Gel, Cellulofine GC-700 (trade name, Chisso
Co., Ltd., particle size 45-100 μm)
And flow water through the column with a peristatic pump.
Through the process, the relationship between the flow rate and the pressure loss ΔP was determined. as a result
Is shown in FIG. As is clear from the figure, it is a soft gel
Agarose gel consolidates above a certain flow rate,
Even if the pressure is increased, the flow rate does not increase.
Hard gels such as cellulose and cellulofine are subject to increased pressure.
The flow rate increases proportionally. Production Example 1 CK Gel A3, a porous cellulose gel (trade name,
Exclusion limit molecular weight of globular protein 5000
100, 100 ml of water and 60 ml of water
And 80 ml of 2M NaOH and stirred at 45 ° C for 1 hour.
Stirred. After stirring, add another 25 ml of epichlorohydrin.
In addition, the mixture was stirred at 45 ° C for 2 hours, and after completion of the reaction, the gel was filtered off.
Wash with water and epoxidize cellulose gel (hereinafter, epoxidized)
Gel). Comparative Production Example 1 To 10 ml of the epoxidized gel obtained in Production Example 1, add sulfani
A solution prepared by dissolving 0.14 g of phosphoric acid in 7.5 ml of water was added.
The pH of the solution was adjusted to 10 by adding 2M NaOH.
After adjusting, it was left at 45 ° C. for 20 hours. After the reaction,
The gel was filtered off and washed with water to fix the sulfanilic acid
I got sougel. Introduced by immobilized sulfanilic acid
The amount of the anionic functional group was 10 μm per 1 ml of the adsorbent.
mol. Comparative Production Example 2 2-aminoethyl hydrogen sulfate instead of sulfanilic acid
Exactly the same as Comparative Production Example 1 except that 0.11 g was used
And the cellulosic acid to which 2-aminoethyl hydrogen sulfate is fixed
I got sougel. 2-aminoethyl hydrogen sulfate fixed
The amount of anionic functional groups introduced by
It was 10 μmol. Comparative Production Example 3 Phosphorylethanolamine instead of sulfanilic acid
Exactly the same as Comparative Production Example 1 except that 0.11 g was used
And a cell in which phosphorylethanolamine is fixed
I got roast gel. Immobilized phosphorylethanolamine
The amount of the anionic functional group introduced by the min was 1 m for the adsorbent.
It was 10 μmol / l. Comparative Production Example 4 1,2-ethanedithiol instead of sulfanilic acid
Exactly the same as Comparative Production Example 1 except that 0.08 g was used.
Then, the cellulos in which 1,2-ethanedithiol is fixed
I got sougel. Immobilized 1,2-ethanedithiol
The amount of anionic functional groups introduced by
It was 10 μmol. Example 1 The molecular weight of about 5 was added to 10 ml of the epoxidized gel obtained in Production Example 1.
Sodium dextran sulfate with a sulfur content of 18%
5 g, and 8 ml of water, and further added 2 M NaOH.
In addition, adjust the pH of the solution to 10 and then at 45 ° C for 17 hours
Left for a while. After completion of the reaction, the gel was filtered and washed with water, and 0.5%
Add ethanolic aqueous solution and leave at room temperature for 20 hours
Then, unreacted epoxy groups were sealed. After the reaction, remove the gel
Dextran sodium sulfate was fixed by filtration and washing
A cellulose gel was obtained. Fixed dextran sulfate
The amount of anionic functional groups introduced from the
It was 29 μmol. Production Example 2 Add 40 ml of the epoxidized gel obtained in Production Example 1
Add 0.5g of amine dissolved in 30ml of water
It was left at 45 ° C. for 20 hours. After completion of the reaction, the gel is filtered and washed with water.
Cellulose gel having amino groups introduced thereinto (hereinafter, amino gel).
No. gel). Comparative Production Example 5 10 ml of water was added to 10 ml of the aminated gel obtained in Production Example 2.
Subsequently, 1 g of glucose 6-barium phosphate was added.
And 0.5 ml of 2M NaOH was added and
It was kept at 5 ° C. NaBH_Four Add 0.1 g at room temperature
It was left for 24 hours. After completion of the reaction, the gel is filtered and washed with water.
Course 6-Cellulose to which phosphoric acid was fixed was obtained. Fixed
Introduced by the added glucose 6-phosphate
The amount of the functional group was 10 μmol / ml of the adsorbent.
Was. Comparative Production Example 6 10 ml of the aminated gel obtained in Production Example 2 was N, N-dimethyl
Suspension in chloroform-formamide (DMF) for a total volume of 18 ml
And 0.27 g of terephthalic acid was dissolved in it
A condensing reagent (N, N'-dicyclohexylcarbodii)
Mido) and stirred at room temperature for 24 hours. After the reaction
The gel is separated by filtration, washed with DMF, ethanol and water in this order.
A cellulose gel with terephthalic acid fixed was obtained. Fixed
Functional group introduced by terephthalic acid introduced
The amount was 10 μmol / ml of the adsorbent. Example 2 Example 10 was added to 10 ml of the aminated gel obtained in Production Example 2.
5 g of dextran sulfate sodium of the same type as used in
And 8 ml of water, and 2 M NaOH 0.5 m
and maintained at 45 ° C. for 1 hour. NaBH_Four
0.1 g was added and left at room temperature for 24 hours. After the reaction
The gel is filtered and washed with water to fix dextran sulfate sodium
The obtained cellulose gel was obtained. Fixed dextran
The amount of anionic functional groups introduced by sulfuric acid is 1 m for the adsorbent.
39 μmol / l. Production Example 3 Porous cellulose gel of the same type as used in Production Example 1
(CK gel A3) 40 ml was suspended in heptane and the whole volume was
To 70 ml. Add 10 ml of 20% NaOH
And nonionic surfactant Tween 20 (trade name, BA
Add 40 drops (Iorad) and shake at 40 ° C for 30 minutes
did. Then add 10 ml of epichlorohydrin and add 40 ml.
Shake at ℃ for 6 hours. After completion of the reaction, the gel was filtered off and ethanol
And then washed with water in order to obtain an epoxidized gel. Example 3 Instead of the epoxidized gel obtained in Production Example 1,
Example 1 except that 10 ml of epoxidized gel was used.
Dextran sulfate sodium
A fixed cellulose gel was obtained. Fixed dext
The amount of anionic functional groups introduced by lansulfuric acid
It was 36 μmol / ml of body. Example 4 Add one end to 10 ml of the epoxidized gel obtained in Production Example 3.
Polyacrylic acid having a mino group (molecular weight: about 1000) 1
g was dissolved in 5 ml of water, and 2M Na was added thereto.
1 ml of OH was added and left at room temperature for 48 hours. End of reaction
After completion, the gel is filtered and washed with water to remove polyacrylic acid
I got Lurose gel. With fixed polyacrylic acid
The amount of the introduced anionic functional group is 5 per ml of the adsorbent.
It was 60 μmol. Poly with amino group at one end
Acrylic acid uses 2-aminoethanethiol as a chain transfer agent
And α, α'-azobisisobutyronitrile (AIB
N) by the low polymerization reaction of acrylic acid with initiator
(The Chemical Society of Japan, 1977, No.
1, pages 88-92, "2-Hydroxyethyl methacrylate"
Synthesis of latet-styrene type ABA type block copolymer
And its structure and wetting ”, Mitsuo Okano, et al.). Example 5 Polyacrylic acid having an amino group at one end (molecular weight of about 1
000) instead of polyacyl having an amino group at one end
Example of manufacture except that 1 g of lylic acid (molecular weight about 10,000) was used
Polyacrylic acid was fixed exactly as in 4.
A cellulose gel was obtained. With fixed polyacrylic acid
The amount of anionic functional groups introduced per 1 ml of adsorbent
It was 5.6 mmol. Has an amino group at one end
Polyacrylic acid is similar to the method described in Example 4
Synthesized according to the method. Production Example 4 To 10 ml of the epoxidized gel obtained in Production Example 3,
A solution prepared by dissolving 0.12 g of amine in 6 ml of water was added.
The aminated gel was obtained in exactly the same manner as in Production Example 2.
Was. Example 6 Obtained in Production Example 4 instead of the aminated gel obtained in Production Example 2
Example 2 except that 10 ml of aminated gel was used.
Dextran sodium sulfate is fixed in exactly the same way
The obtained cellulose gel was obtained. Fixed dextran
The amount of the anionic functional group introduced by the sulfuric acid is the same as that of the adsorbent 1
It was 76 μmol / ml. Production Example 5 CK gel A22 which is a porous cellulose gel (trade name,
Chisso Corporation, exclusion limit molecular weight of globular protein 3000
25, 10 ml of water and 10 ml of water
And 30 ml of 2M NaOH, and shake at 40 ° C. for 20 minutes.
Shaking. Add 12 ml of epichlorohydrin to this and add 4
Shake at 0 ° C. for 3 hours. After completion of the reaction, the gel is filtered and washed with water.
An epoxidized gel was obtained. Production Example 6 CK gel A32 which is a porous cellulose gel (trade name,
Chisso Co., Ltd., exclusion limit molecular weight of globular protein 2000
10,000, particle size 53-125 μm) 25 ml, water 15 ml,
6. 21 ml of 2M NaOH and epichlorohydrin
Epoxidation in the same manner as in Production Example 5 except that 1 ml was used.
I got a gel. Production Example 7 Cellulofine GCL-2, a porous cellulose gel
000m (trade name, manufactured by Chisso Corporation, excluding globular proteins)
Limit molecular weight 3 million, particle size 44-105 μm) 25 m
1, water 25 ml, 2M NaOH 15 ml and epic
Same as Production Example 5 except that 5 ml of lorohydrin was used.
To obtain an epoxidized gel. Production Example 8 Cellulofine GCL-1 which is a porous cellulose gel
000m (trade name, manufactured by Chisso Corporation, excluding globular proteins)
Limiting molecular weight 600,000, particle size 44-105 μm) 25 ml,
25 ml of water, 11.75 ml of 2M NaOH and epi
Same as Production Example 5 except that 4 ml of chlorohydrin was used.
Then, an epoxidized gel was obtained. Production Example 9 Cellulofine GC-70, a porous cellulose gel
0m (trade name, manufactured by Chisso Corporation, exclusion limit of globular proteins)
Molecular weight 400,000, particle size 44-105 μm) 25 ml, water 2
5 ml, 8.75 ml of 2M NaOH and epichloro
The same procedure as in Production Example 5 was repeated except that 3 ml of hydrin was used.
Poxylated gel was obtained. Production Example 10 Cellulofine GC-20, a porous cellulose gel
0m (trade name, manufactured by Chisso Corporation, exclusion limit of globular proteins)
Molecular weight 120,000, particle size 44-105 μm) 25 ml, water 2
5 ml, 7 ml of 2M NaOH and epichlorohydrid
The same procedure as in Production Example 5 was repeated except that 2.5 ml
A xylated gel was obtained. Example 7 Epoxidized gel (CK gel A22) 20 obtained in Production Example 5
ml of the same type of dextra as used in Example 1.
10 g of sodium disulfate and water were added, and the total volume was 33 ml.
And The pH of the solution was adjusted to 10.0 by adding 2M NaOH.
And left at 45 ° C. for 17 hours. After the reaction
The gel is filtered, washed with water, and 0.5% monoethanolamine in water
Add the solution and leave it at room temperature for 20 hours.
Was sealed. After the reaction is completed, the gel is filtered, washed with water, and dextra
A cellulose gel on which sodium disulfate was fixed was obtained.
Anions introduced by immobilized dextran sulfate
The amount of the functional group was 31 μmol / ml of the adsorbent.
Was. Example 8 Epoxidized gel (CK gel A32) 20 obtained in Production Example 6
ml of the same type of dextra as used in Example 1.
10 g of sodium sulfonate and water were added to make the total volume 36 m
1 and the pH of the solution was adjusted to 9.9.
And dextran sulfate
A cellulose gel having sodium acid immobilized thereon was obtained. Fixed
Dextran sulfate introduced anionic agent
The amount of active group was 27 μmol per 1 ml of the adsorbent. Example 9 Epoxidized gel obtained in Production Example 7 (GCL-2000m)
20 ml of the same dexame as used in Example 1
9.3 g of sodium transulfate and water are added to make up the entire volume.
Adjusted to 35 ml and adjusted the pH of the solution to 9.3
Except for the fact that it was the same as in
Obtain a cellulose gel on which sodium lansulfate is fixed.
Was. Anis introduced by immobilized dextran sulfate
The amount of the on-functional group is 30 μmol per 1 ml of the adsorbent.
there were. Example 10 Epoxidized gel obtained in Production Example 8 (GCL-1000m)
20 ml of the same dexame as used in Example 1
9.3 g of sodium transulfate and water are added to make up the entire volume.
Adjusted to 35 ml and adjusted the pH of the solution to 9.3
Except for the fact that it was the same as in
Obtain a cellulose gel on which sodium lansulfate is fixed.
Was. Anis introduced by immobilized dextran sulfate
The amount of the on-functional group is 30 μmol per 1 ml of the adsorbent.
there were. Example 11 Epoxidized gel (GC-700m) 20 obtained in Production Example 9
ml of the same type of dextra as used in Example 1.
9.0 g of sodium disulfate and water were added to make the total volume 36.
ml and the pH of the solution was adjusted to 9.3.
Except that, in the same manner as in Example 7, dextran was used.
A cellulose gel on which sodium sulfate was fixed was obtained. Solid
Anionicity introduced by defined dextran sulfate
The functional group content was 30 μmol / ml of adsorbent.
Was. Example 12 Epoxidized gel (GC-200m) 2 obtained in Production Example 10
In 0 ml, the same type of dextist as used in Example 1
9.0 g of sodium lansulphate and water were added to make a total volume of 3 g.
6 ml, and adjusted the pH of the solution to 9.2.
Except for this, the dextra
A cellulose gel on which sodium disulfate was fixed was obtained.
Anions introduced by immobilized dextran sulfate
The amount of the functional group was 32 μmol per 1 ml of the adsorbent.
Was. Experimental Example 1 and Comparative Experimental Example 1 The absorptions obtained in Examples 1-6 and Comparative Examples 1-6.
The body was washed with a 0.5 M phosphate buffer (pH 7.4).
Then, 0.1 ml of each adsorbent was
Transfer to a tube (1.5 ml capacity)
Acid buffer (pH 7.4) was added to bring the total volume to 1 ml.
0.2 ml of serum containing anti-lipid antibody was added thereto, and 3
Shake at 7 ° C for 2 hours. After this adsorption operation, centrifuge
The gel was allowed to settle, and the anti-lipid antibody titer in the collected supernatant was measured.
It was measured by an enzyme immunoassay (ELISA). Antifat
Quality antibody titers were measured on cardiolipin-coated plates.
Add the diluted sample and perform antigen-antibody reaction.
Add a oxidase-labeled anti-human immunoglobulin antibody,
CS-930 (trade name, Shimadzu Corporation)
Manufactured). Table 1 shows the aniline fixed to each adsorbent.
Name of the compound having an on-functional group and the original blood of each adsorbent
Of anti-lipid antibody titer in the supernatant
Anti-lipid antibody titers are given as percentages as relative antibody titers. From Table 1 it can be seen that dextran sulfate or
Adsorbent with immobilized crylic acid has high ability to adsorb anti-lipid antibody
It turns out that it is excellent. Experimental Example 2 Using the adsorbents obtained in Examples 1 and 7 to 12,
Same as Experimental Example 1 except that the amount of serum added was 0.15 ml.
The relative antibody titer was determined according to the same method. The result
Table 2 shows the names of various water-insoluble porous materials using the fruits.
You. From Table 2, it can be seen that the exclusion limit molecular weight is less than 400,000
Cellulofine GC-700m which is a porous material and cell
Anti-lipid resistance of adsorbent using Lurofine GC-200m
It can be seen that the body adsorption ability is slightly lower. Conversely, the exclusion limit
Anti-lipid antibody absorption even if the molecular weight is too large
It can be seen that the abilities tend to fall. [0069] [Table 1] [0070] [Table 2] [0071] EFFECTS OF THE INVENTION The removal device of the present invention is used to remove anti-lipid antibody from body fluid.
Is selectively removed.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic cross-sectional view of one embodiment of the anti-lipid antibody removing device of the present invention. FIG. 2 is a graph showing the results of examining the relationship between flow rate and pressure loss using three types of gels. [Explanation of Signs] 1 Inlet 2 Outlet 3 Adsorbent 4, 5 Filter 7 Container

Claims (1)

Claims 1. A container having a fluid inlet and a fluid outlet, a fluid and a component contained in the fluid can pass through, but a polyanion compound having a molecular weight of 1,000 or more and having an anionic functional group in a water-insoluble porous body; However, except for DNA, a filter through which the adsorbent of the immobilized anti-lipid antibody cannot pass,
And a device for removing an anti-lipid antibody comprising an adsorbent of the anti-lipid antibody filled in the container. 2. The antimicrobial composition according to claim 1, wherein the anionic functional group comprises at least one selected from the group consisting of a sulfate group, a sulfonic group, a carboxyl group, a thiol group and a phosphate group. A device for removing lipid antibodies. 3. The apparatus for removing an anti-lipid antibody according to claim 1, wherein the water-insoluble porous body comprises a compound having a hydroxyl group.