JPH02180273A - Adsorptive removing agent for beta2-microglobulin - Google Patents
Adsorptive removing agent for beta2-microglobulinInfo
- Publication number
- JPH02180273A JPH02180273A JP63333917A JP33391788A JPH02180273A JP H02180273 A JPH02180273 A JP H02180273A JP 63333917 A JP63333917 A JP 63333917A JP 33391788 A JP33391788 A JP 33391788A JP H02180273 A JPH02180273 A JP H02180273A
- Authority
- JP
- Japan
- Prior art keywords
- microglobulin
- diameter
- adsorption
- beta2
- hydroxyapatite
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 102000015736 beta 2-Microglobulin Human genes 0.000 title abstract description 20
- 108010081355 beta 2-Microglobulin Proteins 0.000 title abstract description 20
- 230000000274 adsorptive effect Effects 0.000 title abstract 3
- 239000002245 particle Substances 0.000 claims abstract description 42
- 229910052588 hydroxylapatite Inorganic materials 0.000 claims abstract description 22
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 claims abstract description 22
- 238000001179 sorption measurement Methods 0.000 claims abstract description 22
- 239000011148 porous material Substances 0.000 claims description 14
- 210000002966 serum Anatomy 0.000 abstract description 9
- 238000000502 dialysis Methods 0.000 abstract description 6
- 206010064553 Dialysis amyloidosis Diseases 0.000 abstract description 4
- 238000005054 agglomeration Methods 0.000 abstract 2
- 230000002776 aggregation Effects 0.000 abstract 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract 1
- 239000003795 chemical substances by application Substances 0.000 description 13
- 238000000034 method Methods 0.000 description 11
- 102000004169 proteins and genes Human genes 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- 239000013078 crystal Substances 0.000 description 6
- 108010088751 Albumins Proteins 0.000 description 4
- 102000009027 Albumins Human genes 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 102100028967 HLA class I histocompatibility antigen, alpha chain G Human genes 0.000 description 3
- 101710197836 HLA class I histocompatibility antigen, alpha chain G Proteins 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000003463 adsorbent Substances 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000010304 firing Methods 0.000 description 2
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 2
- 229910052753 mercury Inorganic materials 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000002002 slurry Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- VZSRBBMJRBPUNF-UHFFFAOYSA-N 2-(2,3-dihydro-1H-inden-2-ylamino)-N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]pyrimidine-5-carboxamide Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C(=O)NCCC(N1CC2=C(CC1)NN=N2)=O VZSRBBMJRBPUNF-UHFFFAOYSA-N 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 101000937544 Homo sapiens Beta-2-microglobulin Proteins 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 239000012888 bovine serum Substances 0.000 description 1
- 238000011437 continuous method Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 102000047279 human B2M Human genes 0.000 description 1
- 230000000951 immunodiffusion Effects 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
Landscapes
- External Artificial Organs (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
Description
【発明の詳細な説明】
［産業上の利用分野］
この発明は、β２−マイクログロブリンの吸着除去剤に
関する。DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to an adsorption and removal agent for β2-microglobulin.
［従来の技術］
長期人工透析患者の合併症の１つである透析アミロイド
症は、患者血液中に存在する低分子量（分子量１１．８
００）のタンパク質であるβ２−マイクログロブリンの
骨組織への蓄積による四肢末梢の疼痛である。[Prior Art] Dialysis amyloidosis, which is one of the complications of long-term dialysis patients, is caused by low molecular weight (molecular weight 11.8
This is pain in the peripheral limbs due to accumulation of β2-microglobulin, a protein of 00), in bone tissue.
透析アミロイド−シス症の要因物質としてβ２−マイク
ログロブリンが確認されたのは比較的近年であり、その
除去法としてろ過透析膜やイオン交換樹脂を用いる方法
が試みられている。しかしながら、従来より提案されて
いる方法では、β２−マイクログロブリンのみを選択的
にを満足できる程度に除去することが困難であり、新し
いβ２−マイクログロブリンの除去吸着剤が望まれてい
る。例えば、特開昭６３−１５９６０号公報によれば。It is relatively recently that β2-microglobulin has been confirmed as a factor in dialysis amyloidosis, and methods using filtration dialysis membranes or ion exchange resins have been attempted as methods for its removal. However, with the conventionally proposed methods, it is difficult to selectively remove only β2-microglobulin to a satisfactory extent, and a new adsorbent for removing β2-microglobulin is desired. For example, according to Japanese Patent Application Laid-Open No. 15960/1983.
β２−マイクログロブリンの吸着除去剤としてプルスハ
イト結晶粒子を出発物質として加熱処理及びアルカリ処
理して得られる結晶粒子を使用することが開示されてい
るが、かかる結晶粒子では分離効果が未だ十分でな（、
粒径もｌＯμｍ程度のものであるから圧損が高（十分な
分離ができない。また、特開昭６３−１５９６１号によ
れば、粒径は、５μｍ〜２０００μｍと太き（、圧損は
解決されたが細孔分布が特定されていないので分離効果
については未だ十分でない。Although it has been disclosed that crystal particles obtained by heating and alkali treatment using purushite crystal particles as a starting material are used as an adsorption/removal agent for β2-microglobulin, the separation effect of such crystal particles is still insufficient ( ,
Since the particle size is about 10 μm, the pressure drop is high (sufficient separation cannot be achieved. Also, according to JP-A-63-15961, the particle size is as large as 5 μm to 2000 μm (the pressure drop has been solved). However, since the pore distribution has not been specified, the separation effect is still insufficient.
［発明が解決しようとする問題点］
従って１本発明の目的は、β２−マイクログロブリンを
選択的に十分に除去することができるβ、−マイクログ
ロブリンの除去吸着剤を提供することである。[Problems to be Solved by the Invention] Accordingly, an object of the present invention is to provide a β,-microglobulin removal adsorbent that can selectively and sufficiently remove β2-microglobulin.
Ｅ問題点を解決するための手段］
本願発明者らは、鋭意研究の結果、特定の粒径、細孔径
及び形状を有するヒドロキシアパタイト粒子集合体を用
いると、β２−マイクログロブリンを選択的に吸着する
ことができることを見出し本発明を完成した。Means for Solving Problem E] As a result of intensive research, the inventors of the present application found that β2-microglobulin can be selectively adsorbed by using a hydroxyapatite particle aggregate having a specific particle size, pore size, and shape. They found that it is possible to do this and completed the present invention.
ヒドロキシアパタイトがタンパク質を吸着する性質は古
くから知られているが、その吸着は、Ｊボタンバク質、
アルブミン等、大分子量タンパク質が中心で、β２−マ
イクログロブリンのような低分子量クンバク質のみを選
択的に吸着することは困難であった１本願発明者らは、
内部に３０ ｎａｒ萌後の細孔を有する球状ヒドロキシ
アパタイト顆粒による分子ふるい効果を利用し、β２−
マイクログロブリンを選択的に吸着させることができる
ことを見出した。The ability of hydroxyapatite to adsorb proteins has been known for a long time.
The inventors of this application found that it was difficult to selectively adsorb only low-molecular-weight proteins such as β2-microglobulin, mainly large-molecular-weight proteins such as albumin.
β2-
It has been found that microglobulin can be selectively adsorbed.
すなわち、本発明は、平均粒径が２０μｍないし５００
０μｍ、内部に直径が３ ｎｍないし２０口ｎｍの連続
細孔を有する実質的に球状のヒドロキシアパタイト粒子
集合体から成るβ２−マイクログロブノンの吸着除去剤
を提供する。That is, in the present invention, the average particle size is 20 μm to 500 μm.
Provided is an adsorption/removal agent for β2-microglobunon consisting of a substantially spherical hydroxyapatite particle aggregate having a diameter of 0 μm and internal continuous pores of 3 nm to 20 nm in diameter.
［発明の効果］
本発明により、β２−マイクログロブリンを選択的に満
足できる程度に除去することができる。[Effects of the Invention] According to the present invention, β2-microglobulin can be selectively removed to a satisfactory extent.
従って、本発明の８２−マイクログロブリン吸着除去剤
に人工透析時の患者血清を通過させ、血清中の８２−マ
イクログロブリンを吸着除去することができ、それによ
って人工透析患者の透析アミロイド−シス症を軽減する
ことができる。Therefore, by passing patient serum during artificial dialysis through the 82-microglobulin adsorption/removal agent of the present invention, 82-microglobulin in the serum can be adsorbed and removed, thereby preventing dialysis amyloidosis in artificial dialysis patients. It can be reduced.
［発明の詳細な説明］
本発明のβ２−マイクログロブリン吸着除去剤を構成す
るヒドロキシアパタイト粒子集合体の平均粒径は２０μ
自ないし５０００μ園、好ましくは１１００ｔｉないし
５０［１ｕｍである。平均粒径が２０１１ｆｆｉよりも
小さいとカラムでの使用時に閉塞が生じ易（、また、逆
に平均粒径が５０００ＬＬｍよりも大きいとβ２−マイ
クログロブリンの吸着能力が低下する。平均粒径は、試
料の顕微鏡写真を撮りその写真より粒径を測定すること
により測定することができる。[Detailed Description of the Invention] The average particle size of the hydroxyapatite particle aggregate constituting the β2-microglobulin adsorption/removal agent of the present invention is 20μ
The thickness is from self to 5000μ, preferably from 1100ti to 50[1um]. If the average particle size is smaller than 2011ffi, clogging is likely to occur during column use (on the other hand, if the average particle size is larger than 5000LLm, the adsorption ability of β2-microglobulin will be reduced. It can be measured by taking a microscopic photograph of the particles and measuring the particle size from the photograph.
本発明の吸着除去剤を構成するヒドロキシアパタイト粒
子は、内部に直径ｌ口ｎｌｌないし２００ｎｍ、好まし
くは２０ ｎｍないし５０ ｎｍの連続細孔を有する。The hydroxyapatite particles constituting the adsorption removal agent of the present invention have continuous pores with a diameter of 1 to 200 nm, preferably 20 to 50 nm.
連続細孔径が上記範囲よりも大きくても小さくてもβ２
−マイクログロブリンを選択的に吸着除去することがで
きない、なお２粒子の細孔径は特願昭６３−５８０５９
号に示すような水銀正大法により測定することができる
。Even if the continuous pore diameter is larger or smaller than the above range, β2
- Microglobulin cannot be selectively adsorbed and removed, and the pore diameter of the two particles is determined by patent application No. 63-58059.
It can be measured by the mercury Seitai method as shown in the issue.
また、本発明の吸着除去剤を構成するヒドロキシアパタ
イト粒子の細孔容積は５０％以上であることが好ましく
、８０％以上であることがさらに好ましい、細孔容積が
５０％未満ではβ２−マイクログロブリンの吸着能力が
低下する。Further, the pore volume of the hydroxyapatite particles constituting the adsorption removal agent of the present invention is preferably 50% or more, more preferably 80% or more, and if the pore volume is less than 50%, β2-microglobulin adsorption capacity decreases.
本発明の吸着除去剤を構成するヒドロキシアパタイト粒
子は実質的に球状である１粒子の形状が球状からはずれ
ると充填率が低下し、かつ、均一な充填が困難となり吸
着力に影響を及ぼす。The hydroxyapatite particles constituting the adsorption/removal agent of the present invention are substantially spherical, and if the shape of one particle deviates from the spherical shape, the filling rate decreases and uniform filling becomes difficult, which affects the adsorption force.
本発明の吸着除去剤を構成するヒドロキシアパタイト粒
子集合体は、例えば、特願昭６３−５８０５９号に記載
された方法により製造することができる。すなわち、従
来より周知の方法により製造された針状ヒドロキシアパ
タイト結晶のスラリーを、特定条件下で撹拌することに
より、球状ヒドロキシアパタイトが得られ、さらに必要
に応じて焼成することにより製造することができる。す
なわち、ヒドロキシアパタイト結晶のスラリー（１０〜
６０重量％）を密閉容器内において回転翼により１０〜
３Ｈｒｐｍの回転数により１時間ないし２０時間撹拌を
行ない、（得られた球状のヒドロキシアパタイト粒子を
１００℃ないし９００℃で焼成することにより（得るこ
とができる。The hydroxyapatite particle aggregate constituting the adsorption-removal agent of the present invention can be produced, for example, by the method described in Japanese Patent Application No. 63-58059. That is, spherical hydroxyapatite is obtained by stirring a slurry of acicular hydroxyapatite crystals produced by a conventionally known method under specific conditions, and can be further produced by firing if necessary. . That is, a slurry of hydroxyapatite crystals (10~
60% by weight) in a sealed container using a rotary blade for 10~
The mixture can be obtained by stirring at a rotational speed of 3 Hrpm for 1 to 20 hours and firing the obtained spherical hydroxyapatite particles at 100 to 900°C.
本発明の８２−マイクログロブリン吸着除去剤は、例え
ばこれをカラム又はシリンジ等の容器に充填し、人工透
析時の患者血清を通過させて使用することができる。こ
の際、カラムに充填する吸着除去剤の量は、特に限定さ
れないが、通常１０ ｇないし１ｏｏｏ ｇ、好ましく
は３００ｇないし５００ｇであり、流通させる患者血清
の流量は通常１Ｏｎｌ／分ないし１００ ｍ１７分、好
ましくは１０ ｍ１７分ないし３０ ｍ１７分である。The 82-microglobulin adsorption/removal agent of the present invention can be used, for example, by filling it into a container such as a column or a syringe, and allowing patient serum during artificial dialysis to pass through the container. At this time, the amount of adsorption removal agent packed into the column is not particularly limited, but is usually 10 g to 100 g, preferably 300 g to 500 g, and the flow rate of patient serum to be distributed is usually 1 Onl/min to 100 m17 min. Preferably it is 10 m17 minutes to 30 m17 minutes.
以下、本発明を実施例に基づきより詳細に説明する。も
っとも、本発明は下記実施例に限定されるものではない
。Hereinafter, the present invention will be explained in more detail based on Examples. However, the present invention is not limited to the following examples.
［実施例１
１ｉ■」
ヒドロキシアパタイト粒子集合体の製造ヒドロキシアパ
タイト結晶の２８重量％スラノーを回転翼により８０
ｒｐｎ＋の回転数で８時間撹拌し１球状のヒドロキシア
パタイト粒子集合体を得た。さらに、同粒子を電気炉を
用いて５００℃で３時間焼結を行なった。[Example 1 1i■] Production of hydroxyapatite particle aggregate A 28% by weight slano of hydroxyapatite crystals was heated to 80% by weight using a rotary blade.
The mixture was stirred at a rotation speed of rpn+ for 8 hours to obtain a spherical hydroxyapatite particle aggregate. Furthermore, the same particles were sintered at 500° C. for 3 hours using an electric furnace.
得られたヒドロキシアパタイト粒子集合体の平均粒径及
び細孔径を測定したところ、平均粒径は２００μｍない
し５００μ閣、細孔径は２０％ｍないし３０％ｍであっ
た。平均粒径及び細孔径の測定は具体的には以下のよう
にして行なった。すなわち、平均粒径はミクロメーター
を組み込んだ顕微鏡により測定した。また、細孔径は水
銀圧入ボリシメーター［Ｍｉｃｒｏｍｅｒｉｔｉｃｓ
Ａｕｔｏｐｏｒｅ ９２００．島津製作所製］により測
定した。また、得られたヒドロキシアパタイト粒子集合
体を顕微鏡にて観察したところ、各粒子は実質的に球状
であった。When the average particle size and pore size of the obtained hydroxyapatite particle aggregate were measured, the average particle size was 200 μm to 500 μm, and the pore size was 20% m to 30% m. Specifically, the average particle size and pore size were measured as follows. That is, the average particle size was measured using a microscope equipped with a micrometer. In addition, the pore diameter can be measured using a mercury intrusion volisimeter [Micromeritics].
Autopore 9200. [manufactured by Shimadzu Corporation]. Furthermore, when the obtained hydroxyapatite particle aggregate was observed under a microscope, each particle was found to be substantially spherical.
１五±１
β、−マイクログロブリンの吸着除去（バッチ法）ヒト
β２−マイクログロブリン（尿由来）０．１５ｍｇをウ
シ血清３．０ ｌｌ１ｌに溶解して約５０ ｐｐｍ相当
の模擬患者血清を調製し、１０ｍ１三角フラスコ内で、
実施例１で得られたヒドロキシアパタイト粒子１．Ｏｒ
ｎｌ （約０．６５ ｇｌ と共にマグネット回転子に
よるスターラー攪拌（約２０Ｏｒｐｍｌを９０分間行な
った。その後、０２−マイクログロブリンを常法である
５ＲＩＤ法（抗β２−マイクログロブリンヤギ血清によ
る一次免疫拡散法）によりＢ２−マイクログロブリンの
量を定量したところ、約７０％のβ２マイクログロブリ
ンの吸着効果が見られた。また、同時に実施したアルブ
ミン及び総タンパク質の吸着状況はそれぞれ１８％、２
８％であった。なお、総タンパク質は屈折計法により、
アルブミンはＢＣＧ法により定量した。Adsorption and removal of 15±1 β,-microglobulin (batch method) 0.15 mg of human β2-microglobulin (derived from urine) was dissolved in 3.0 liters of bovine serum to prepare a simulated patient serum equivalent to approximately 50 ppm. , in a 10m Erlenmeyer flask,
Hydroxyapatite particles obtained in Example 1 1. Or
nl (approximately 0.65 gl and stirred with a magnetic rotor for 90 minutes (approximately 20 Orpml) for 90 minutes. After that, 02-microglobulin was subjected to the conventional 5RID method (primary immunodiffusion method using anti-β2-microglobulin goat serum). When the amount of B2-microglobulin was quantified, approximately 70% of the adsorption effect of β2-microglobulin was observed.Also, the adsorption status of albumin and total protein, which was carried out at the same time, was 18% and 2-microglobulin, respectively.
It was 8%. In addition, total protein was determined by refractometer method.
Albumin was quantified by the BCG method.
これらの結果から、本発明のＢ２−マイクログロブリン
吸着除去剤は、Ｂ２−マイクログロブリンを選択的に吸
着することがわかる。These results show that the B2-microglobulin adsorption/removal agent of the present invention selectively adsorbs B2-microglobulin.
実」１１ユ
Ｂ２−マイクログロブリンの吸着除去（連続法）実施例
１で得られたヒドロキシアバタイ［−粒子０．６５ｇを
１．０ ｍｌシリンジに充填したカラムに実施例２で調
製した模擬患者血清を、マイクロフィーダーを用い、
０．０８ ｃｃ／＋＊ｉｎ、の割合で通過させた６通過
時間と、シリンジを通過した後の模擬血清中の６２−マ
イクログロブリン量、総タンパク質量及びアルブミン量
を１５分間隔で測定した。Adsorption and removal of microglobulin (continuous method) The simulated patient prepared in Example 2 was placed in a column filled with 0.65 g of hydroxyabatai particles obtained in Example 1 in a 1.0 ml syringe. Serum, using a microfeeder,
The amount of 62-microglobulin, total protein, and albumin in the simulated serum after passing through the syringe was measured at 15-minute intervals over a 6-passage time at a rate of 0.08 cc/+*in.
結果を下記表に示す。The results are shown in the table below.
これらの結果から、本発明のβ２−マイクログロブリン
吸着除去剤は、β２−マイクログロブリンを選択的に吸
着することがわかる。These results show that the β2-microglobulin adsorption/removal agent of the present invention selectively adsorbs β2-microglobulin.
Claims (2)
直径が３ｎｍないし２００ｎｍの連続細孔を有する実質
的に球状のヒドロキシアパタイト粒子集合体から成るβ
＿２−マイクログロブリンの吸着除去剤。(1) β consisting of substantially spherical hydroxyapatite particle aggregates with an average particle size of 20 μm to 5000 μm and internal continuous pores with a diameter of 3 nm to 200 nm.
_2-Adsorption and removal agent for microglobulin.
請求項１記載の吸着除去剤。(2) The adsorption removal agent according to claim 1, wherein the pores have a diameter of 20 nm to 50 nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63333917A JPH02180273A (en) | 1988-12-31 | 1988-12-31 | Adsorptive removing agent for beta2-microglobulin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63333917A JPH02180273A (en) | 1988-12-31 | 1988-12-31 | Adsorptive removing agent for beta2-microglobulin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH02180273A true JPH02180273A (en) | 1990-07-13 |
Family
ID=18271411
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63333917A Pending JPH02180273A (en) | 1988-12-31 | 1988-12-31 | Adsorptive removing agent for beta2-microglobulin |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH02180273A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6878269B2 (en) | 1996-01-31 | 2005-04-12 | Kaneka Corporation | Device for body fluid purification and system for body fluid purification |
-
1988
- 1988-12-31 JP JP63333917A patent/JPH02180273A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6878269B2 (en) | 1996-01-31 | 2005-04-12 | Kaneka Corporation | Device for body fluid purification and system for body fluid purification |
| US7279106B2 (en) | 1996-01-31 | 2007-10-09 | Kaneka Corporation | Adsorbent and method for adsorbing a chemokine in body fluid |
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