JPS58166263A - Preparation of standard apoprotein a-i - Google Patents
Preparation of standard apoprotein a-iInfo
- Publication number
- JPS58166263A JPS58166263A JP4816382A JP4816382A JPS58166263A JP S58166263 A JPS58166263 A JP S58166263A JP 4816382 A JP4816382 A JP 4816382A JP 4816382 A JP4816382 A JP 4816382A JP S58166263 A JPS58166263 A JP S58166263A
- Authority
- JP
- Japan
- Prior art keywords
- apoprotein
- blood
- serum
- column
- standard
- 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.)
- Granted
Links
- 102000006410 Apoproteins Human genes 0.000 title claims abstract description 39
- 108010083590 Apoproteins Proteins 0.000 title claims abstract description 39
- 238000002360 preparation method Methods 0.000 title claims description 3
- 210000002966 serum Anatomy 0.000 claims abstract description 31
- 210000004369 blood Anatomy 0.000 claims abstract description 16
- 239000008280 blood Substances 0.000 claims abstract description 16
- 239000011347 resin Substances 0.000 claims abstract description 7
- 229920005989 resin Polymers 0.000 claims abstract description 7
- 210000002381 plasma Anatomy 0.000 claims abstract 4
- 102000015779 HDL Lipoproteins Human genes 0.000 claims description 8
- 108010010234 HDL Lipoproteins Proteins 0.000 claims description 8
- 238000004062 sedimentation Methods 0.000 abstract description 12
- 230000002159 abnormal effect Effects 0.000 abstract description 3
- 241000283707 Capra Species 0.000 abstract description 2
- 230000005484 gravity Effects 0.000 abstract description 2
- 230000003053 immunization Effects 0.000 abstract description 2
- 230000037356 lipid metabolism Effects 0.000 abstract description 2
- 210000002826 placenta Anatomy 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 239000003381 stabilizer Substances 0.000 abstract description 2
- 102000004895 Lipoproteins Human genes 0.000 abstract 1
- 108090001030 Lipoproteins Proteins 0.000 abstract 1
- 101100041681 Takifugu rubripes sand gene Proteins 0.000 abstract 1
- 230000003100 immobilizing effect Effects 0.000 abstract 1
- 239000000463 material Substances 0.000 abstract 1
- 239000004576 sand Substances 0.000 abstract 1
- 238000000034 method Methods 0.000 description 17
- 102000005666 Apolipoprotein A-I Human genes 0.000 description 5
- 108010059886 Apolipoprotein A-I Proteins 0.000 description 5
- 229920002684 Sepharose Polymers 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 235000004252 protein component Nutrition 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 2
- ATDGTVJJHBUTRL-UHFFFAOYSA-N cyanogen bromide Chemical compound BrC#N ATDGTVJJHBUTRL-UHFFFAOYSA-N 0.000 description 2
- 238000011033 desalting Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 238000003018 immunoassay Methods 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 108010056308 A-1 antigen Proteins 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- 206010008909 Chronic Hepatitis Diseases 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 101100055841 Danio rerio apoa1 gene Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 102000014190 Phosphatidylcholine-sterol O-acyltransferase Human genes 0.000 description 1
- 108010011964 Phosphatidylcholine-sterol O-acyltransferase Proteins 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000005238 degreasing Methods 0.000 description 1
- 230000015961 delipidation Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 230000000951 immunodiffusion Effects 0.000 description 1
- 238000000760 immunoelectrophoresis Methods 0.000 description 1
- 238000001155 isoelectric focusing Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000003169 placental effect Effects 0.000 description 1
- 229910000027 potassium carbonate Inorganic materials 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000003127 radioimmunoassay Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M sodium chloride Inorganic materials [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/96—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood or serum control standard
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Hematology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
【発明の詳細な説明】
本発明は高比重リポ蛋白（以下Ｈ１）Ｌと略す）から遊
離したアポ蛋白Ａ−１（以下遊離アポＡ−１と言う）と
ＨＤ Ｌ顆粒表面に存在するアポ蛋白Ａ−１（以下ＨＤ
Ｌ・アポＡ−１と言う）の分離法即ちアポ蛋白Ａ−１濃
度測定用標準品に関するものである。Detailed Description of the Invention The present invention relates to apoprotein A-1 (hereinafter referred to as free apoA-1) released from high-density lipoprotein (hereinafter referred to as H1L) and apoprotein present on the surface of HDL granules. A-1 (hereinafter HD
This paper relates to a standard product for measuring apoprotein A-1 concentration, that is, a method for separating L. apoA-1).
リボ蛋白は脂質と蛋白質成分より構成されており、その
蛋白質はアポ蛋白と呼ばれている。Riboproteins are composed of lipid and protein components, and the protein is called apoprotein.
アポ蛋白には多くの種類があるが、アポ蛋白Ａ−１はこ
れらアポ蛋白の中で血清又は血漿中でその濃度が最も高
いことが知られている。アポ蛋白Ａ−１は主としてＨＤ
Ｌ顆粒表面に存在し。Although there are many types of apoproteins, apoprotein A-1 is known to have the highest concentration in serum or plasma among these apoproteins. Apoprotein A-1 is mainly found in HD
Exists on the surface of L granules.
ＨＤＬの可溶化に重要な働きをしているほか。In addition, it plays an important role in solubilizing HDL.
レシチンコレステロールアシルトランスフェラーゼの活
性化作用を有している。It has an activating effect on lecithin cholesterol acyltransferase.
最近、アポ蛋白の測定が注目されてきており。Recently, the measurement of apoprotein has been attracting attention.
特にアポ蛋白Ａ−１及びＡ−１の測定は脂質代謝異常の
鑑別また慢性肝炎及び肝硬変の重症度の判定に有効であ
ることが知られている。In particular, measurement of apoprotein A-1 and A-1 is known to be effective in differentiating lipid metabolism abnormalities and determining the severity of chronic hepatitis and liver cirrhosis.
アポ蛋白Ａ−１の測定法には免疫学的測定法として一元
放射状免疫拡散法（以下５ＲＩＤ法と略す）、免疫電気
泳動法、オフタロニー法。Immunoassay methods for measuring apoprotein A-1 include one-way radial immunodiffusion method (hereinafter abbreviated as 5RID method), immunoelectrophoresis method, and ophthalmology method.
ラジオイムノアッセイ法、エンザイムイムノアッセイ法
、電気化学的性質を利用した等電点電気泳動法、８ＤＳ
ゲル電気泳動法等が使用されている。特に５ＲＩＤ法、
免疫電気法−動法は測定感度にすぐれ且つ操作の簡便さ
から日常検査に適した測定法として普及が期待されてい
る。Radioimmunoassay method, enzyme immunoassay method, isoelectric focusing method using electrochemical properties, 8DS
Gel electrophoresis and the like are used. Especially the 5RID method,
The immunoelectrical method-dynamic method is expected to become popular as a measuring method suitable for daily testing because of its excellent measurement sensitivity and ease of operation.
しかし、これらの方法を使用する場合標準品として精製
したアポ蛋白Ａ−［を用いるが、精製アポ蛋白Ａ−１は
繁雑な操作と特殊な装置を使用し脱脂操作を行なう等手
間がかかる。しかも脱脂操作が行なわれているので、血
清又は血漿中のアポ蛋白とは挙動が異なることが予想さ
れ好ましくない。一方正常新鮮血清又は血漿を１００％
として順次希釈し検量線を作成する方法があるが、新鮮
な血清又は血漿を使用しないと二重の沈降線あるいは不
鮮明な沈降線を生じる。However, when these methods are used, purified apoprotein A-[ is used as a standard product, but purified apoprotein A-1 requires complicated operations and a degreasing operation using special equipment, which is time-consuming. Moreover, since a delipidation operation has been performed, it is expected that the behavior will be different from that of apoprotein in serum or plasma, which is not preferable. On the other hand, 100% normal fresh serum or plasma
There is a method of creating a calibration curve by serially diluting the sample, but if fresh serum or plasma is not used, a double sedimentation line or an unclear sedimentation line will result.
この原因は採取した血清又は血漿中のアポ蛋白・ Ａ−
１が時間の経過と伴にＨＤＬ・アポＡ−１と遊離アポＡ
−１に分離し混在してくるためである。この様な血清又
は血漿を標準品として使用すると正確な測定値が得られ
にくい。従ってこれらの現象を避けるには新鮮血清又は
血漿を標準品として使用する必要１が、する。しかし測
定毎に正常新鮮血清又は血漿を標準品として使用するこ
とは通常不可能である。The cause of this is apoprotein A- in the collected serum or plasma.
1 becomes HDL/apoA-1 and free apoA over time.
This is because they are separated into −1 and mixed together. When such serum or plasma is used as a standard product, it is difficult to obtain accurate measured values. Therefore, to avoid these phenomena, it is necessary to use fresh serum or plasma as a standard. However, it is usually impossible to use normal fresh serum or plasma as a standard for each measurement.
なぜならば検査室に集まる血清又は血漿は異常もしくは
異常と思われるものがほとんどであるため、標準品とし
て使用するには好ましくない。そこで新鮮血清又は血漿
と同等に正確に測定し得る標準品即ち、二重の沈降線あ
るいは不鮮明な沈降線を生じない標準血清が要望されて
いた。This is because most of the serum or plasma collected in testing laboratories is abnormal or appears to be abnormal, so it is not desirable to use it as a standard product. Therefore, there has been a need for a standard serum that can be measured as accurately as fresh serum or plasma, that is, a standard serum that does not produce double sedimentation lines or unclear sedimentation lines.
本発明者は二重の沈降線あるいは不鮮明な沈降線を生じ
る原因となっている遊離アポＡ−１を除去するため超遠
心分離法あるいはゲルロ過法等種々試みたが除去し得な
かった。しかし。The present inventor attempted various methods such as ultracentrifugation and gel filtration in order to remove free apoA-1, which causes double sedimentation lines or indistinct sedimentation lines, but was unable to remove it. but.
さらに鋭意研究を進めた結果１１ Ｄ Ｌ・アポＡ−１
及び遊離アポＡ−１は抗アポ蛋白Ａ−１抗体に対する親
和性に明らかに差があることを見出し、ＨＤＬ・アポＡ
−１及び遊離アポＡ−１を選択的に分離し得る方法を完
成した。As a result of further intensive research 11 D L Apo A-1
We found that there was a clear difference in the affinity of anti-apoprotein A-1 antibodies between HDL and free apoA-1.
We have completed a method that can selectively separate ApoA-1 and free ApoA-1.
本発明は多量に入手可能な血液即ち胎盤後血あるいは有
効期限切れの輸血用血液を原料として、これら血液の血
清成分または血液をそのまま抗アポ蛋白Ａ−［抗体カラ
ムに接触させることを特徴とする遊離アポＡ−１分離法
即ち標準品の調製法である。The present invention uses blood that is available in large quantities, that is, postplacental blood or expired blood for transfusion, as a raw material, and the serum components of these blood or blood are directly brought into contact with an anti-apoprotein A-[antibody column. This is the ApoA-1 separation method, ie, the standard preparation method.
次に本発明について詳細に説明する。Next, the present invention will be explained in detail.
先ず胎盤後血または有効期限切れの輸血用血液を比重１
．２０に調製した後、遠心分離を行ない上部画分を捕集
する。次いで上部画分は脱塩操作を行なう。First, placental blood or expired blood for transfusion is diluted with a specific gravity of 1.
．． 20, centrifugation is performed and the upper fraction is collected. The upper fraction is then subjected to a desalting operation.
一方、セファローズ４Ｂ（ファルマシア社発売）にアポ
蛋白特異抗体を固定化させた抗アポ蛋白Ａ−１抗体カラ
ムを調製し、緩衝液で平衡化しカラムに充填しておく。On the other hand, an anti-apoprotein A-1 antibody column in which an apoprotein-specific antibody is immobilized on Sepharose 4B (manufactured by Pharmacia) is prepared, equilibrated with a buffer solution, and packed into the column.
該カラムに脱塩操作を行なった後のアポ蛋白Ａ−１を含
む画分を浸透させ緩衝液で流出させると最初に目的物で
あるＨＤＬ・アポ蛋白Ａ−１を含む画分が流出して来る
。この自分に安定化剤を加え凍結乾燥することによりア
ポ蛋白Ａ−１濃度測定用標準血清が得られる。When the fraction containing apoprotein A-1 after desalting is permeated into the column and eluted with a buffer solution, the fraction containing HDL/apoprotein A-1, which is the target product, flows out first. come. By adding a stabilizer to this serum and lyophilizing it, a standard serum for measuring apoprotein A-1 concentration can be obtained.
なお、抗アポ蛋白Ａ−１抗体カラムは以下の方法で調製
できる。Incidentally, an anti-apoprotein A-1 antibody column can be prepared by the following method.
例えば単離精製したアポ蛋白Ａ−１を動物（山羊、家兎
等）に免疫して得られた抗血清をそのまま次の操作に使
用してもよいが、好ましくは得られた抗血清からさらに
アポ蛋白Ａ−１特異抗体を得９次の操作に使用するのが
よい。For example, an antiserum obtained by immunizing an animal (goat, rabbit, etc.) with isolated and purified apoprotein A-1 may be used as is in the next operation, but it is preferable to further use the obtained antiserum. It is preferable to obtain an apoprotein A-1 specific antibody and use it in the ninth step.
特異抗体を得る方法としてはアポ蛋白Ａ−１抗原を、セ
ファロース４Ｂ等の不溶性樹脂に固定化させカラムに充
填し、緩衝液で平衡化させた後、諌カラム番こ抗血清を
浸透させた後常法の手段で処理すればアポ蛋白Ａ−１特
異抗体が得られる。この特異抗体を活性化させた不溶性
樹脂例えばセファロース４Ｂに接触させればセファロー
ス４Ｂｊこ特異抗体が固定化される。The method for obtaining specific antibodies is to immobilize the apoprotein A-1 antigen on an insoluble resin such as Sepharose 4B, fill it in a column, equilibrate it with a buffer solution, and then infiltrate the Isa column with Banko antiserum. Apoprotein A-1 specific antibodies can be obtained by treatment using conventional methods. When this specific antibody is brought into contact with an activated insoluble resin such as Sepharose 4B, the specific antibody is immobilized on Sepharose 4Bj.
次に参考例、実施例で本発明を説明する。Next, the present invention will be explained with reference examples and examples.
参考例 アポ蛋白Ａ−１抗体カラム
セファロース４Ｂ ８．９を２モル炭酸カリウム５１Ｉ
／に懸濁し、氷冷後１ ｒｔｒｌのアセトニトリルに２
１の臭化シアンを溶解した溶液０．２５ｍ４を加え数分
間攪拌した後グラスフィルター（８Ｇ）で口過し、残渣
を１０倍量の蒸留水、０，１モル炭酸水素ナトリウムで
洗浄する。得られた残渣はアポ蛋白特異抗原８０■をＯ
１モル炭酸水素ナトリウム７１Ｒ１に溶解したものに加
え。Reference example Apoprotein A-1 antibody column Sepharose 4B 8.9 to 2 mol potassium carbonate 51I
/, cooled on ice, and diluted with 1 rtrl of acetonitrile.
Add 0.25 m4 of a solution in which cyanogen bromide (1) was dissolved, stir for several minutes, filter through a glass filter (8G), and wash the residue with 10 times the amount of distilled water and 0.1 mol sodium bicarbonate. The obtained residue contains 80 μg of apoprotein-specific antigen.
Add to that dissolved in 1M sodium bicarbonate 71R1.
４℃以下で１６時間反応させる。反応後口過し残渣を蒸
留水で洗浄後０．０２％アジ化ナトリウムを含む０．５
モル塩化す） ＩＪウムに懸濁させ、４℃以下で保存す
る。React at 4°C or lower for 16 hours. After the reaction, the residue was filtered and washed with distilled water.
(Molar chloride) Suspend in IJum and store at below 4°C.
実施例
胎盤後面ｌｌに乾燥した臭化カリウム２９１．６ｇを加
え混合した後、４℃で４４００Ｏｒｐｍ、 ４８時間
遠心分離した。分離後遠心管上部画分を捕集し、脱脂綿
を用いて不溶物を日別した。Example 291.6 g of dried potassium bromide was added to the posterior surface of the placenta 11 and mixed, followed by centrifugation at 4400 rpm at 4° C. for 48 hours. After separation, the upper fraction of the centrifuge tube was collected, and insoluble matter was separated using absorbent cotton.
ロ別後０．２モルショ糖−塩化す） ＩＪウム溶液に平
衡化したセファデックスＧ−２５（ファルマシア社発売
）を充填したカラムに浸透させ同溶液で流出させた。流
出液は各５縦に分画し連続的に蛋白成分を分析しアポ蛋
白Ａ−ｌを含む両分を得た。次いでこの両分をまとめ０
１２モルショ糖−塩化ナトリウム溶液に平衡化したアポ
蛋白Ａ−１抗体カラムに浸透させ同溶液にて流出させた
。流出液は各７履ｌに分画し、連続的に蛋白成分を分析
し、）（ＤＩ、・アポＡ−１を含む画分を捕集した。After separation, the column was permeated into a column filled with Sephadex G-25 (manufactured by Pharmacia) equilibrated with a 0.2M sucrose-chloride solution, and the same solution was allowed to flow out. The effluent was fractionated vertically into 5 sections and the protein components were continuously analyzed to obtain both fractions containing apoprotein A-1. Next, combine these two parts and 0
It was permeated into an apoprotein A-1 antibody column equilibrated with a 12M sucrose-sodium chloride solution and eluted with the same solution. The effluent was fractionated into 7 liter portions, continuously analyzed for protein components, and fractions containing )(DI, ApoA-1) were collected.
得られたＨＤＬ・アポＡ−（を含む両分をバイアルビン
に０．５ ｄずつ分注し凍結乾燥し血中アポ蛋白Ａ−１
濃度測定用標準血清を得たＯ
この標準血清１バイアルに対し生理食塩液０．５ｄに溶
解し、標準血清試料とした。Both the obtained HDL and apoA-(containing) were dispensed into vials in 0.5 d portions and lyophilized to obtain blood apoprotein A-1.
A standard serum for concentration measurement was obtained. One vial of this standard serum was dissolved in 0.5 d of physiological saline to prepare a standard serum sample.
正常人新鮮血清及び４℃で７日間放置した正常人の血清
をそれぞれ生理食塩液にて１０倍に希釈し試料とした。Fresh serum from a normal person and serum from a normal person left at 4°C for 7 days were each diluted 10 times with physiological saline and used as samples.
標準血清試料及び試料を８ＲＩＤ法用に調製した抗アポ
蛋白Ａ−１抗血清を含むアガロース板上に各５μｌ添加
し室温にて４８時間水平台上で静置し沈降線を比較観察
した。標準血清及び新鮮血清では一本の鮮明な沈降線を
得たが、４℃で７日間放置した血清では二重の沈降線を
得た。5 μl of each of the standard serum sample and the sample were added to an agarose plate containing anti-apoprotein A-1 antiserum prepared for the 8RID method, and the plate was left standing on a horizontal stand at room temperature for 48 hours, and the sedimentation lines were comparatively observed. A single clear sedimentation line was obtained with the standard serum and fresh serum, but a double sedimentation line was obtained with the serum that had been left at 4°C for 7 days.
Claims (1)
れた後、血清、血漿又は血液を該不溶性樹脂に接触させ
高比重リポ蛋白から遊離したアポ蛋白Ａ−１を血清、血
漿又は血液から分離させることを特徴とするアポ蛋白Ａ
−１標準品の調製法After the apoprotein A-1 antibody is immobilized on an insoluble resin and equilibrated, serum, plasma, or blood is brought into contact with the insoluble resin to remove apoprotein A-1 released from high-density lipoproteins from the serum, plasma, or blood. Apoprotein A characterized by being separated
-1 Preparation method of standard product
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4816382A JPS58166263A (en) | 1982-03-27 | 1982-03-27 | Preparation of standard apoprotein a-i |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4816382A JPS58166263A (en) | 1982-03-27 | 1982-03-27 | Preparation of standard apoprotein a-i |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS58166263A true JPS58166263A (en) | 1983-10-01 |
| JPH0474670B2 JPH0474670B2 (en) | 1992-11-26 |
Family
ID=12795709
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP4816382A Granted JPS58166263A (en) | 1982-03-27 | 1982-03-27 | Preparation of standard apoprotein a-i |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS58166263A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS60253871A (en) * | 1984-05-30 | 1985-12-14 | Nippon Koutai Kenkyusho:Kk | Anti-apo a-1 antibody |
| JPS6264956A (en) * | 1985-09-18 | 1987-03-24 | Teijin Ltd | Method for detecting lung surface active substance and reagent kit therefor |
| JPS63501037A (en) * | 1985-09-27 | 1988-04-14 | フア−マシア・ア−・ベ− | Method for measuring lipoproteins and apolipoproteins |
| US5070270A (en) * | 1987-03-04 | 1991-12-03 | Mitsubishi Denki K.K. | Brush device |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0688614B2 (en) * | 1992-07-13 | 1994-11-09 | 静男 佐藤 | Paper protector having multiple structure and manufacturing apparatus thereof |
-
1982
- 1982-03-27 JP JP4816382A patent/JPS58166263A/en active Granted
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS60253871A (en) * | 1984-05-30 | 1985-12-14 | Nippon Koutai Kenkyusho:Kk | Anti-apo a-1 antibody |
| JPH0453516B2 (en) * | 1984-05-30 | 1992-08-26 | Nippon Kotai Kenkyusho Kk | |
| JPS6264956A (en) * | 1985-09-18 | 1987-03-24 | Teijin Ltd | Method for detecting lung surface active substance and reagent kit therefor |
| JPS63501037A (en) * | 1985-09-27 | 1988-04-14 | フア−マシア・ア−・ベ− | Method for measuring lipoproteins and apolipoproteins |
| US5070270A (en) * | 1987-03-04 | 1991-12-03 | Mitsubishi Denki K.K. | Brush device |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0474670B2 (en) | 1992-11-26 |
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