JPS5861466A - Measuring method serumnological agglutination reaction - Google Patents

Measuring method serumnological agglutination reaction

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Publication number
JPS5861466A
JPS5861466A JP15952881A JP15952881A JPS5861466A JP S5861466 A JPS5861466 A JP S5861466A JP 15952881 A JP15952881 A JP 15952881A JP 15952881 A JP15952881 A JP 15952881A JP S5861466 A JPS5861466 A JP S5861466A
Authority
JP
Japan
Prior art keywords
protein
methylated
carrier
hapten
latex
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
Application number
JP15952881A
Other languages
Japanese (ja)
Other versions
JPH0153424B2 (en
Inventor
Hiroko Koyanagi
小柳　裕子
Susumu Sofue
晋 祖父江
Iwao Sozaki
曽根崎　巖
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
YATORON KK
Mitsubishi Kagaku Iatron Inc
Original Assignee
YATORON KK
Mitsubishi Kagaku Iatron Inc
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Application filed by YATORON KK, Mitsubishi Kagaku Iatron Inc filed Critical YATORON KK
Priority to JP15952881A priority Critical patent/JPS5861466A/en
Publication of JPS5861466A publication Critical patent/JPS5861466A/en
Publication of JPH0153424B2 publication Critical patent/JPH0153424B2/ja
Granted legal-status Critical Current

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    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding

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  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Analytical Chemistry (AREA)
  • Biotechnology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)

Abstract

PURPOSE:To provide a method for adsorbing and binding hapten which is non- protein extremely easily on a carrier by interposing methylated protein between the non-protein haptens and the carrier. CONSTITUTION:Methylated protein is mixed and prepd. with a suitable equiv. of non-protein hapten and a carrier, whereby the bound material of the non-protein hapten-carrier is easily obtained. Here the carrier refers to the latex and immobilized blood corpuscles prepd. by known methods. The reasons why the methylated protein is effective as interposing protein lie in the probability that when protein is methylated (+) changes are increased and hydrophobicity is intensified on account of which the adsorbing and binding between the methylated protein and the non-protein hapten and between the methylated protein and the carrier take place satisfactorily. Thus non-protein antigens such as glycolipids, polysaccharides, nucleic acids, and their separated materials are adsorbed easily on the carriers without degrading the activity of the antigens.

Description

【発明の詳細な説明】 本発明は非蛋白性物質を抗原として担体に吸着させ、免
疫化学的に凝集反応を起させる関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to adsorbing a non-protein substance as an antigen to a carrier and immunochemically causing an agglutination reaction.

従来、血液、尿、その他体液中に存在する生理活性物質
を測定する方法として、その物質のもつ免疫学的活性を
利用した免疫化学的方法が用いられている。すなわち抗
原と抗体とを反応させて生じる凝集反応もしくは凝集阻
止反応によって、該抗原もしくは該抗体を測定する方法
である。Conventionally, as a method for measuring physiologically active substances present in blood, urine, and other body fluids, an immunochemical method that utilizes the immunological activity of the substance has been used. That is, it is a method of measuring an antigen or an antibody by an agglutination reaction or an agglutination inhibition reaction generated by reacting the antigen with the antibody.

その際の反応を測定し易くする為に、一般に微細粒子を
抗原もしくは抗体の担体として用いることが行われてい
る。担体にはラテックス、血球、活性炭、、ベントナイ
ト、カオリン、細菌粒子などが用いられる。これらの中
でラテックスおよび固定化血球が最も一般に用いられて
いる。In order to make it easier to measure the reaction at that time, fine particles are generally used as carriers for antigens or antibodies. Latex, blood cells, activated carbon, bentonite, kaolin, bacterial particles, etc. are used as carriers. Of these, latex and immobilized blood cells are the most commonly used.

これらの担体に抗原もしくは抗体を吸着あるいは共有結
合させて用いるが、一般には吸着が簡単であり、かつ抗
原活性あるいは抗体活性をそこなうことがないので繁用
される。。These carriers are used by adsorbing or covalently bonding antigens or antibodies to them, and they are commonly used because adsorption is easy and does not impair antigen or antibody activity. .

担体に抗原もしくは抗体を吸着させるＫはそれらが蛋白
質であることが好都合であった。It is advantageous for K to adsorb antigens or antibodies to carriers to be proteins.

抗体は蛋白質そのものであるが、抗原の場合でも蛋白質
の場合の方がよく成功し又いる。Antibodies are proteins themselves, but even in the case of antigens, the use of proteins has been more successful.

担体特にラテックスあるいは血球に吸着するには吸着力
が働かなければならないのであるが、ラテックスあるい
は血球と蛋白質との間の吸着力が一番強い条件が見い出
さ４利用されているのである。In order to adsorb to a carrier, especially latex or blood cells, adsorption force must work, and the conditions under which the adsorption force between latex or blood cells and proteins is the strongest have been discovered and utilized4.

非蛋白質抗原を相体に吸着させるには該抗原独自の吸着
条件を見出すか、又は抗原を免疫学的反応に関わらない
蛋白質と共有結合して、該蛋白質を介して抗原な担体に
吸着させる方法がある。In order to adsorb a non-protein antigen to a phase, it is necessary to find adsorption conditions unique to the antigen, or to covalently bond the antigen to a protein that is not involved in the immunological reaction, and then adsorb the antigen to a carrier via the protein. There is.

抗原独自の吸着条件を見出す方法は非常に繁雑であり必
ずしも他の場合に応用できない。The method of finding adsorption conditions unique to an antigen is extremely complicated and cannot necessarily be applied to other cases.

また、免疫学的反応に関わらない蛋白質と共有結合させ
た上で担体に吸着させる方法は他の場合にも応用できる
けれども、これは抗原活性を損わない共有結合方法を見
出すのが困難である。Additionally, although the method of covalently bonding proteins that are not involved in immunological reactions and then adsorbing them to a carrier can be applied to other cases, it is difficult to find a covalent bonding method that does not impair antigen activity. .

本発明は抗原活性を損わないで簡単に蛋白質と非蛋白質
抗原とを吸着させた上で該蛋白質を介し℃、非蛋白質抗
原な担体に吸着させる方法及びその方法にする試薬を提
供するものである。The present invention provides a method for easily adsorbing a protein and a non-protein antigen without impairing antigen activity, and then adsorbing the non-protein antigen to a non-protein antigen carrier via the protein, and a reagent for the method. be.

脂質、糖質、核酸等の非蛋白質のものは一般にハシテン
と呼ばれ、その物単独では動物に注射しても抗体を作ら
せる能力、すなわち抗原性が極めて弱いかほとんど無い
。Non-protein substances such as lipids, carbohydrates, and nucleic acids are generally referred to as hasitenes, and when injected into animals alone, they have very weak or almost no antigenicity, i.e., the ability to produce antibodies.

プロティンと呼ぶ。It's called protein.

従って、ハプテンもしくは−・ブテン様物質にキャリア
・プロティンを結合あせる方法は、単に吸着結合のみな
らず、共有結合等に於けるまで従来からよく検討され利
用されている。Therefore, methods for bonding carrier proteins to hapten- or butene-like substances have been well studied and utilized in the past, including not only adsorption bonding but also covalent bonding and the like.

しかしながら前述したように、／・ブテン蛋白質を共有
結合させる場合はともすると抗原性が低下するか或は抗
原性が変ってしまうことが応々にしである。その為に抗
原性が低下しないか或は変ってしまわない結合方法が求
められる。However, as mentioned above, when a butene protein is covalently bonded, the antigenicity often decreases or changes. Therefore, a binding method that does not reduce or change antigenicity is required.

また＃に述ぺたようにハプテンは抗原性が不完全なもの
であるが、特に脂質の場合、例］えばホスファチツルセ
リンやカルジオＩＪピンはそれぞれレシチンおよびコレ
ステロールとの共存ミセルな人工的に作って抗原と′ｆ
るといった如く複雑なものもある。In addition, as mentioned in #, haptens have incomplete antigenicity, but especially in the case of lipids, for example, phosphatitulserin and cardio-IJ pin are artificially produced micelles that coexist with lecithin and cholesterol, respectively. The antigen and 'f
There are some complicated ones, such as

担体にバッテンを結合させる方法に於ても数多くのもの
が試みられて（・る力１、簡便で確かな方法が求められ
る。）・ブテン例え＆ｆ月旨實等を相体に簡便にして確
かな方法で吸着結合させることができれば液体中の抗体
グ）１を６Ａ１［定−′ｆろのに便利である。カルヅオ
ＩＪピン（丁梅毒トレポネーマ感染による抗体を、ＤＮ
Ａｔ了自己免疫疾患である全身性工１３テマトーデスで
見られろ抗ＤＮＡ抗体を検出することカーできる。また
さらに細菌等微生物細胞壁多糖体は該微生物による抗体
を検出すること力１できる。Many methods have been tried for bonding battens to carriers (a simple and reliable method is needed). If adsorption bonding can be carried out using a suitable method, it would be convenient to filtrate the antibody G)1 in the liquid. Calduo IJ pin (DN) to detect antibodies caused by Treponema pallidum infection
At present, it is possible to detect anti-DNA antibodies seen in the autoimmune disease systemic 13-thematosus. Furthermore, cell wall polysaccharides of microorganisms such as bacteria can be used to detect antibodies produced by the microorganisms.

一般に、担体に抗原または抗体カー非特異的に吸着結合
するには担体のもつ電荷と抗原または抗体のもつ電荷と
か作用し合って（・る。Generally, in order for an antigen or antibody to nonspecifically adsorb and bond to a carrier, the charges on the carrier interact with the charges on the antigen or antibody.

従って担体に抗原または抗体を吸着結合するには、その
時に用いられる担体と吸着させる抗原または抗体のそれ
ぞれの持つ電荷に最適の条件のρＢが選ばれるのが望ま
しい。通常は特定のｐＨの緩衝液が用いられる。それで
もなおハプテン特に非蛋白性／′−ゾテンは担体に吸着
結合させるのが難しい。Therefore, in order to adsorb and bond an antigen or antibody to a carrier, it is desirable to select the optimal condition ρB for the charge possessed by the carrier used at that time and the antigen or antibody to be adsorbed. Usually, a buffer solution with a specific pH is used. Nevertheless, haptens, especially non-proteinaceous/'-zothenes, are difficult to adsorb and bond to carriers.

本発明ではメチル化蛋白質を非蛋白性］・ゾテ／と担体
の間に介在させることで結果的に非蛋白性ハプテンを担
体に吸着結合させる方法を提供する。非蛋白性ノ・ブテ
ンと担体との間に介在するものは、非蛋白性／・ブテン
と吸着結合し、かつまた担体と吸着結合するものであれ
ば用いられる。数ある蛋白質のなかにはそのままでも介
在蛋白質の役割をはたすものもあるだろうが、そｊを探
すのは一般的でない。そこで本発明者らは通常入手可能
な蛋白質を用いて介在蛋白質とする方法を創意工夫した
。The present invention provides a method in which a methylated protein is interposed between a non-protein hapten and a carrier, thereby adsorbing and bonding a non-protein hapten to the carrier. The substance interposed between the non-protein butene and the carrier may be used as long as it binds to the non-protein butene by adsorption and also binds to the carrier by adsorption. Although there are many proteins that may play the role of intervening proteins as they are, it is not common to search for them. Therefore, the present inventors devised a method for using commonly available proteins as intervening proteins.

通常蛋白質と呼ばれるものには豆類等穀物から得られる
他物蛋白質や動物の血漿や卵等から得られる動物蛋白質
や細菌体から得られる細菌蛋白質が挙げられる。What is usually called protein includes foreign proteins obtained from grains such as beans, animal proteins obtained from animal plasma, eggs, etc., and bacterial proteins obtained from bacterial bodies.

本発明にはこれらの蛋白質はいずれも用いることが可能
である。通常容易に人手できる蛋白質を用いるのが便利
である。ウシ血清アルブミンや卵白アルブミン等が容易
に入手できて便利であるが、本発明ではこれらに限らず
メチル化できる蛋白質ならば使用可能である。Any of these proteins can be used in the present invention. It is usually convenient to use a protein that can be easily prepared manually. Bovine serum albumin, ovalbumin, etc. are easily available and convenient, but the present invention is not limited to these, and any protein that can be methylated can be used.

蛋白質は大きな分子であり、分子の中に数多くのカルボ
キシル基（−Ｃ０ＯＨｌ　　とアミノ＆　（−ＮＨ、ｌ
　　を持っているが、このカルボキシル基をメチル基和
することで蛋白質はメチル化される。もちろん、カルボ
キシル基以外の部分がメチル化されても良い、介在蛋白
質としてメチル化蛋白質が有効である理由は充分には研
究さねではいないが、蛋白質がメチル化されることによ
って（＋）向電を多くシ、かつ疎水性が高まるので、メ
チル化蛋白質と非蛋白性ハプテンの間およびメチル化蛋
白質と担体の間の吸着結合がうまくゆくものらしい。Proteins are large molecules with many carboxyl groups (-C0OHl and amino & (-NH, l).
Proteins are methylated by adding a methyl group to this carboxyl group. Of course, moieties other than carboxyl groups may be methylated, and the reason why methylated proteins are effective as intervening proteins has not been fully researched, but methylation of proteins can lead to (+) It seems that adsorption bonds between methylated proteins and non-protein haptens and between methylated proteins and carriers are likely to be successful because of the increased hydrophobicity and increased hydrophobicity.

メチル化蛋白質を適当量の非蛋白性ハシテンおよび担体
と混合調整することによって、簡便に非蛋白性ハゲテン
−担体連結物が得られる。望ましくは担体にメチル化蛋
白質をあらかじめ吸着結合した後に非蛋白性ハシテンを
吸着結合するのがよい。By mixing and preparing a methylated protein with an appropriate amount of non-protein halogen and a carrier, a non-protein halogen-carrier linkage can be easily obtained. Preferably, the methylated protein is adsorbed and bonded to the carrier in advance, and then the non-proteinaceous protein is adsorbed and bonded to the carrier.

ここで担体とは公知の方法によって作製されろラテック
スおよび固定化血球を言う。なおラテックスなる名称は
一般に使用されるポリスチレンラテックスのみならず、
他のポリマー或は種々のコポリマーラテックスが該当す
る。Here, the carrier refers to latex and immobilized blood cells prepared by a known method. The name latex refers not only to the commonly used polystyrene latex, but also to
Other polymers or various copolymer latexes are suitable.

次に実施例を挙げて説明する。Next, an example will be given and explained.

実施例１ メチル化蛋白質の作製 ■　１２のウシ血清アルブミン（Ｂ８Ａ）を１００−の
無水メタノールに懸濁する。これに濃塩酸帆８４−を攪
拌しながら滴下する。室温暗所でときどき攪拌しなから
１日１、ｌヒ、好ましくは３日間放音する。その後遠心
して蛋白沈殿を集め、これを無水メタノール、次いでエ
ーテル中で、洗絨が黄色に着色しなくなるまで洗う。こ
れを真空ｒジケータ中で乾燥保存する。これをメチル化
ウシ血清アルブミン（’Ｍ−ＢＳＡＩと言う。使用時１
幅水溶液とする。Example 1 Preparation of methylated protein ■ 12 bovine serum albumin (B8A) is suspended in 100-g absolute methanol. Concentrated hydrochloric acid 84- is added dropwise to this while stirring. Leave the mixture at room temperature in a dark place with occasional stirring and emit sound once a day, preferably for 3 days. Thereafter, the protein precipitate is collected by centrifugation and washed in anhydrous methanol and then in ether until the washed cellulose is no longer colored yellow. This is stored dry in a vacuum radiator. This is called methylated bovine serum albumin ('M-BSAI).
Make it into an aqueous solution.

（２）蛋白質として卵白アルブミン（ＯＶＡＩとした以
外はＢＳＡの場合に′！ｌＡじてメチル化する。これを
メチル化卵白アルブミン（Ｍ−ＯＶＡ）と言う。(2) Except for ovalbumin (OVAI), the protein is methylated as '!lA in the case of BSA. This is called methylated ovalbumin (M-OVA).

■　蛋白質としてウマ血清ガンマグロブリン（ＨｏＳＧ
Ｉとした外はＢＳＡの場合に命じてメチル化する。これ
をメチル化ウマ血清がンマグロプリン（Ｍ−Ｈｏ　Ｓ　
Ｇ　）と言う。■ Horse serum gamma globulin (HoSG) is a protein.
Except for I, methylation is performed in the case of BSA. This is converted into methylated horse serum, which is converted into maglopurine (M-HoS).
G).

実施例２ メチル化蛋白質吸着ラテックスの作製 ■　ポリスチレンラテックスの５憾懸濁欣をあらかじめ
適当な緩衝ｇに透析しておく。Example 2 Preparation of methylated protein adsorption latex ■ Five suspensions of polystyrene latex were dialyzed in advance against an appropriate buffer.

別に用意した１％ＭＢ８Ａを３〜ｉｏ容好ｉ　Ｌ　＜　
ハ４〜７容と５憾ラデツクス懸濁液とを混合する。この
混合液を３７℃で１時間ときどき攪拌しながらインキュ
ベートする。この懸濁液を８．０００　）、ｐ、ｍ、で
１５分間遠心分離してラテックスをＥｏｌ収する。ラテ
ックスを緩衝液で均一に懸濁しこれなさらに遠心分離し
てラテックスを回収する。Separately prepared 1% MB8A was mixed with 3 ~ io good i L <
C. Mix 4 to 7 volumes of Radex suspension. This mixture is incubated at 37° C. for 1 hour with occasional stirring. This suspension is centrifuged at 8,000 p, m for 15 minutes to collect the latex. The latex is uniformly suspended in a buffer solution and further centrifuged to recover the latex.

回収したラテックスを緩衝液で均一にほぐし１０鴫懸濁
液とする。The collected latex is uniformly loosened with a buffer solution to form a 10-ml suspension.

Ｃ４ＭＢＳＡをＭＯＶＡとした外は■と同様にする。The procedure is the same as ■ except that C4MBSA is MOVA.

■ノ　ＭＢＳＡをＭ−ＨｏＳＧとした外は■と同様にす
る、 実施例３ メチル化蛋白質吸着ラテックスに脂質を吸着コレステロ
ール３０〜１５０■、好マしくは７０〜１００ｗｇ、ウ
シ心臓力ルノオリビン１〜６η、好マシクは２〜４■、
および卵黄レシチン１０〜７０ｗｇ、好ましくは２０〜
５０■をエタノール１０ｍ１Ｋ浴解する。このエタノー
ル＃沿を緩衝液９０＋ｄＫ添加混合して懸濁液を作り脂
質抗原とする。■No Same as ■ except that M-HoSG was used instead of MBSA. Example 3 Adsorbing lipids to methylated protein adsorption latex Cholesterol 30-150■, preferably 70-100wg, Bovine heart power Lunoolivine 1-6η , Good Masik is 2~4■,
and egg yolk lecithin 10-70wg, preferably 20-70wg
50 μm was dissolved in 10 ml of 1K bath of ethanol. This ethanol mixture is mixed with buffer solution 90+dK to make a suspension and used as a lipid antigen.

実施例２で作製したメチル化蛋白質吸着ラテックスのｌ
θ％懸濁＊１ｇｖｃ上記脂質抗原液を３０−〜１００ｍ
、好ましくは５０−〜７０−を加えて静かに混合する。1 of the methylated protein adsorption latex prepared in Example 2
θ% suspension *1 gvc of the above lipid antigen solution from 30 to 100 m
, preferably from 50 to 70, and mix gently.

これを４〜８℃で５時間以上、好ましくは２４時間とき
どき攪拌しながら放置する。これを遠心分離してラテッ
クスを回収して緩衝液にラテックスが０．５優となるよ
うに均一に懸濁する。This is left at 4-8° C. for 5 hours or more, preferably 24 hours with occasional stirring. This is centrifuged to collect the latex, and the latex is uniformly suspended in a buffer solution to a concentration of 0.5%.

ここで作られた試薬は梅毒陽性血清の検出としてスライ
ド凝集反応用試薬に適する。これを従来公知の炭末凝集
反応試薬と比較すると反応が早く出る点で、スクリーニ
ングに好適である。The reagent prepared here is suitable as a slide agglutination reaction reagent for detecting syphilis-positive serum. Comparing this with conventionally known charcoal powder aggregation reaction reagents, it is suitable for screening because it reacts quickly.

ヒト血清と反応させた結果の一部を第１ｇＫ示した。Part of the results of the reaction with human serum is shown for the first gK.

実施例４ ＭＢＳＡの代りにＭＯＶＡあるいは Ｍ　Ｈｏ　Ｓ　Ｇ　Ｌだ外は実施例３と同様に行う。Example 4 MOVA or instead of MBSA The same procedure as in Example 3 was carried out except for M　Ho　S　G　L.

実施例５ ポリスチレンラテックスの代りに比重の大きいラテック
スを用いる。伊１えばメタクリレートラテックスを用い
た外は実施例３と同様に行う。Example 5 Latex with high specific gravity is used instead of polystyrene latex. Example 1 The same procedure as in Example 3 was carried out except that methacrylate latex was used.

ここで作られた試薬はマイクロタイタープレート用凝集
反応試薬に好適である。ヒト血清と反応させた結果の一
部を第２表に示した。The reagent prepared here is suitable as an agglutination reaction reagent for microtiter plates. Table 2 shows some of the results of the reaction with human serum.

実施例６ メチル化蛋白質吸着血球 担体として、公知の方法で作製した２、５チ固定化動物
血球を用いた外は実施例２に準じて行う。Example 6 The procedure of Example 2 was repeated, except that 2 or 5 immobilized animal blood cells prepared by a known method were used as the methylated protein-adsorbed hemocyte carrier.

実施例７ メチル化蛋白質吸着ラテックスの代りにメチル化蛋白質
吸着血球を用いた外は実施例３に準じて行う。Example 7 The procedure of Example 3 was repeated except that methylated protein-adsorbed hemocytes were used instead of the methylated protein-adsorbed latex.

ここで作られた試薬はマイクロタイターゾレート凝集反
応およびスライド凝集〜応の両方に好適である。The reagents made here are suitable for both microtitrezolate agglutination reactions and slide agglutination reactions.

ヒト血清と反応させた結果の一部を表３に示す。Table 3 shows some of the results of the reaction with human serum.

実施ｆ１１８ 公知の方法で仔牛胸腺から精製し７４　Ｄ　Ｎ　Ａを５
００μｔ　／　ｍｌの濃度としｌ　Ｑ　（１’Ｃ１１０
分間加熱急冷する。これを非、蛋白質抗原とした外は実
施ｆｌ＋　３．４．５．７に準じて行う。Implementation f118 74 DNA was purified from calf thymus using a known method.
Let the concentration be 00 μt/ml and l Q (1'C110
Heat and quench for a minute. The procedure was carried out in accordance with Example fl+ 3.4.5.7, except that this was used as a non-protein antigen.

ここで作られた試薬は全身性エリテマトーデスなど自己
免疫疾患に現われる抗ＤＮＡ抗体の検出に用いられる、 実施例９ 細菌、酵母、カビ等の微生物細胞壁多糖体を公知の方法
で抽出精製してこれを非蛋白質抗原とした外は実施？１
１３．４．５．７に準じて行う。The reagent produced here is used to detect anti-DNA antibodies that appear in autoimmune diseases such as systemic lupus erythematosus. Is it implemented except for non-protein antigens? 1
13.4.5.7.

ここで作られた試薬は該微生物が感染した場合あるいは
該微生物基しくは該微生物細胞壁多糖体を免疫して得た
抗血清の検査に用いられる。The reagent prepared here is used for testing antisera obtained when the microorganism is infected or by immunization with the microorganism base or cell wall polysaccharide.

表１ Ａ：本発明によるＭＢＳＡを用いた方法Ｂ：ＢｓＡを用
いた方法 表２ Ａ：ＭＢＳＡを用いた時 Ｂ：　　Ｉ　　用いない時 表３ Ａ：本発明による血球凝集法−スライド凝集法 Ｂ：　　　　　　　　　　　　−マイクロタイター凝集
法 Ｃ：ＭＢＳＡを用いない血球凝集法−スライド凝集法Table 1 A: Method using MBSA according to the present invention B: Method using BsA Table 2 A: When using MBSA B: When not using I Table 3 A: Hemagglutination method according to the present invention - slide agglutination method B: - Microtiter agglutination method C: Hemagglutination method without MBSA - Slide agglutination method

Claims (4)

【特許請求の範囲】[Claims] （１）　　非蛋白質抗原をメチル化蛋白質を介して担体
に吸着させる事を峙徴とす“る免疫学的測定方法。(1) An immunoassay method that uses the adsorption of a non-protein antigen to a carrier via a methylated protein as a defining characteristic. （２）　　メチル化蛋白質がメチル化アルブミンである
第１項記載の方法。(2) The method according to item 1, wherein the methylated protein is methylated albumin. （３）　　非蛋白質抗原が脂質、糖脂質、多糖類、核酸
、およびそれらの分解物または結合物あるいは混合物で
ある第１項記載の方法。(3) The method according to item 1, wherein the non-protein antigen is a lipid, a glycolipid, a polysaccharide, a nucleic acid, or a decomposition product, a conjugate, or a mixture thereof. （４）担体がラテックス粒子および血球である第１項記
載の方法。(4) The method according to item 1, wherein the carrier is latex particles and blood cells.
JP15952881A 1981-10-08 1981-10-08 Measuring method serumnological agglutination reaction Granted JPS5861466A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP15952881A JPS5861466A (en) 1981-10-08 1981-10-08 Measuring method serumnological agglutination reaction

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
JP15952881A JPS5861466A (en) 1981-10-08 1981-10-08 Measuring method serumnological agglutination reaction

Publications (2)

Publication Number Publication Date
JPS5861466A true JPS5861466A (en) 1983-04-12
JPH0153424B2 JPH0153424B2 (en) 1989-11-14

Family

ID=15695730

Family Applications (1)

Application Number Title Priority Date Filing Date
JP15952881A Granted JPS5861466A (en) 1981-10-08 1981-10-08 Measuring method serumnological agglutination reaction

Country Status (1)

Country Link
JP (1) JPS5861466A (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS63142268A (en) * 1986-11-26 1988-06-14 ベーリンガー・マンハイム・ゲゼルシヤフト・ミツト・ベシユレンクテル・ハフツング Carrying material and manufacture thereof
JPH01209370A (en) * 1987-12-24 1989-08-23 Boehringer Mannheim Gmbh Method and apparatus for measuring immunologically active substance

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS63142268A (en) * 1986-11-26 1988-06-14 ベーリンガー・マンハイム・ゲゼルシヤフト・ミツト・ベシユレンクテル・ハフツング Carrying material and manufacture thereof
JPH01209370A (en) * 1987-12-24 1989-08-23 Boehringer Mannheim Gmbh Method and apparatus for measuring immunologically active substance

Also Published As

Publication number Publication date
JPH0153424B2 (en) 1989-11-14

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