JPS6141966A - 免疫学的測定法 - Google Patents

免疫学的測定法

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Publication number
JPS6141966A
JPS6141966A JP16500885A JP16500885A JPS6141966A JP S6141966 A JPS6141966 A JP S6141966A JP 16500885 A JP16500885 A JP 16500885A JP 16500885 A JP16500885 A JP 16500885A JP S6141966 A JPS6141966 A JP S6141966A
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JP
Japan
Prior art keywords
measurement
antibody
radiation
antigen
measurement container
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
JP16500885A
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English (en)
Other versions
JP2584611B2 (ja
Inventor
ジユハニ　イー．アイ．ルオトラ
ハンヌ　ハルジユンマー
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.)
Thermo Fisher Scientific Oy
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Labsystems Oy
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Publication date
Application filed by Labsystems Oy filed Critical Labsystems Oy
Publication of JPS6141966A publication Critical patent/JPS6141966A/ja
Application granted granted Critical
Publication of JP2584611B2 publication Critical patent/JP2584611B2/ja
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

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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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G01N33/54326—Magnetic particles
    • 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
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S436/00—Chemistry: analytical and immunological testing
    • Y10S436/806—Electrical property or magnetic property
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S436/00—Chemistry: analytical and immunological testing
    • Y10S436/824—Immunological separation techniques

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Food Science & Technology (AREA)
  • General Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Biotechnology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Pathology (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Investigating Or Analyzing Materials By The Use Of Magnetic Means (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

(57)【要約】本公報は電子出願前の出願データであるた
め要約のデータは記録されません。

Description

【発明の詳細な説明】 本発明は固体物質の微小球上に付着させた蛍光又は燐光
トレーサーを用いて抗体又は抗原を標識した、蛍光又は
燐光による免疫測定法に関するものである。
固相蛍光による免疫測定法（固相蛍光免疫測定法）にお
いて、抗原又は抗体はしばしばポリスチレン又はポリア
クリリックで作られた微小球上に付着されている。この
方法では、更に大きな表面を用いることも可能であり、
該表面には抗体又は抗原も又より付着し易い、固体表面
に付着した抗原又は抗体は、測定されるべき試料中に存
在する抗体又は抗原及び蛍光分子で標識した抗体又は抗
原と各々反応する。試料中に存在する抗体又は抗原が多
いほど、固相に付着する標識物質が少なくなる。液相中
に残る抗体又は抗原を分離し、次いで固相に付着してい
る標識物質の量を蛍光計を用いて測定することにより、
試料中の抗体又は抗原の濃度を算出する。
先行技術の方法においては、余分に標識された抗体又は
抗原は蛍光測定の前に洗浄除去しなければならない。
特定の放射免疫測定法（ＲＩ　Ａ）においては、液体の
除去又は洗浄の間磁場を用いることによりその位置を保
つ、磁性物質を含む微小球が使用される。
アメリカ合衆国特許明細書第４４３８０８８号（第６欄
５第８行ないし第１４行）には又、固相として磁性粒子
を使用する方法が開示されている。
競合反応の後、粒子は磁石を用いて沈澱され、次いで上
澄液中に残った標識粒子の放射線が試験管の壁を通して
測定される。開示された測定法によると、結合標識の量
は微小部分を沈澱させた後、反応混合物から間接的に測
定される。
それ故、測定精度は良くない、液相の測定も又、更に難
かしい、すなわち、測定を妨げたり、又は誤差を生じさ
せる物質が存在する可能性がある。
本発明の固相蛍光免疫測定法においては、磁性微小球を
固相として使用する０反応後、磁性微小球を反応容器の
壁面に引きつけ、次いで蛍光（又は燐光）放射線を壁を
通して測定する。
本発明の目的は、蛍光による測定前に余分のトレーサー
を微小球を含む測定容器の外に除去する必要のない蛍光
固相免疫測定法を提供することにある。
本発明の方法において、測定容器を空にして微小球を洗
浄する操作は省略することができる。それ故、測定にお
ける液体の移動は液体の添加のみである。これにより測
定はより迅速となり、その自動化が確実に促進される。
しかしながら、測定精度は良好であり、モして液相は測
定を損わないであろう。
いっそう好ましい実施態様としては、励起又は放射波長
の波長において強く吸収する着色剤を加えるとよい、こ
の結果、液相からの背景放射の影響は全く除かれる。
本発明の方法は、例えば以下のように行う：測定される
べき抗体の抗原を磁性物質を含む大きさ約０．２ないし
１０ｐＬｍのプラスチック微小球に付着させる。このプ
ラスチックは、例えばポリスチレン又はポリアクリリッ
クであり、そして磁性物質は例えば鉄、コバルト又はニ
ッケルである。
微小球を透明な測定容器例えば試験管中に入れ、この中
に測定されるべき抗体を含む試料及び蛍光分子を用いて
標識した抗体を加える０反応の完結後、励起放射線を試
料に照射するとともに蛍光放射線を測定容器の底部を通
して検出器に集めることにより、固相上の抗体に付着し
た標識抗体の量を蛍光計を用いて測定する。
更に、蛍光計に磁場を生じる手段を備え、そしてこれに
よって微小球を測定時間中測定容器の底部に引きつける
。
更に、測定前に、着色剤を粒状又は溶解状態で試料に加
える。この着色剤は励起放射線又は放出放射線の波長で
強い吸収を示す、こり方法によって、微小球の表面に付
着したトレーサーの量を、液相中に残った過剰のトレー
サーによる干渉又は測定時の背景放射による干渉なしで
正確に測定することができる０着色剤は好ましくは黒色
物例えばすすである。
勿論、着色剤を、抗原と抗体との反応に干渉しないよう
に、又はトレーサーの蛍光を妨げないように選択すべき
である。
本方法は燐光を用いる方法にも又適用できることは勿論
である。
特許出願人　ラブシステムズ　オイ （ほか１名）

Claims (6)

    【特許請求の範囲】
  1. （１）抗原又は抗体を磁性粒子に付着させ、測定の間該
    粒子を磁場を用いて測定容器の壁面に引きよせ、励起放
    射線を磁性粒子に照射し、そして蛍光又は燐光放射線を
    該測定容器の壁面を通して検出器に集めることを特徴と
    す る、抗原又は抗体を固体粒子に付着させ、該抗原又は抗
    体を測定容器中で測定されるべき試料中に存在する抗体
    又は抗原及び蛍光物質で標識した抗体又は抗原と各々反
    応させ、その後該固体粒子に付着したトレーサーを適す
    る放射線によって励起し、次いで放出される蛍光又は燐
    光放射線を測定する免疫学的測定法。
  2. （２）抗原又は抗体を、磁性物質を加えたポリマー粒子
    に付着させることを特徴とする、特許請求の範囲第１項
    記載の方法。
  3. （３）測定前に、励起放射線の波長で強い吸収を示す着
    色剤を測定容器中に加えることを特徴とする、特許請求
    の範囲第１項又は第２項記載の方法。
  4. （４）測定前に、放出放射線の波長で強い吸収を示す着
    色剤を測定容器中に加えることを特徴とする、特許請求
    の範囲第１項ないし第３項のうちいずれか１項記載の方
    法。
  5. （５）着色剤が黒色物である、特許請求の範囲第３項又
    は第４項記載の方法。
  6. （６）励起放射線を試料に照射し、次いで蛍光又は燐光
    放射線を測定容器の底部を通して検出器に集めることを
    特徴とする、特許請求の範囲第１項ないし第５項のうち
    いずれか１項記載の方法。
JP60165008A 1984-07-26 1985-07-25 免疫学的測定法 Expired - Lifetime JP2584611B2 (ja)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FI842992A FI842992A0 (fi) 1984-07-26 1984-07-26 Immunologiskt definitionsfoerfarande.
FI842992 1984-07-26

Publications (2)

Publication Number Publication Date
JPS6141966A true JPS6141966A (ja) 1986-02-28
JP2584611B2 JP2584611B2 (ja) 1997-02-26

Family

ID=8519415

Family Applications (1)

Application Number Title Priority Date Filing Date
JP60165008A Expired - Lifetime JP2584611B2 (ja) 1984-07-26 1985-07-25 免疫学的測定法

Country Status (6)

Country Link
US (1) US4731337A (ja)
EP (1) EP0169434B1 (ja)
JP (1) JP2584611B2 (ja)
AT (1) ATE67858T1 (ja)
DE (1) DE3584196D1 (ja)
FI (1) FI842992A0 (ja)

Cited By (6)

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JPS63108265A (ja) * 1986-10-25 1988-05-13 Nippon Telegr & Teleph Corp <Ntt> レ−ザ磁気免疫測定方法
JPH02151766A (ja) * 1988-12-02 1990-06-11 Nippon Telegr & Teleph Corp <Ntt> レーザ磁気免疫測定法
JPH07159407A (ja) * 1993-12-06 1995-06-23 Olympus Optical Co Ltd 光学的免疫測定方法及びこれに用いられる免疫測定装置
JP2010256116A (ja) * 2009-04-23 2010-11-11 Nippon Koden Corp 尿中バイオマーカ測定方法及び測定システム
JP2017510810A (ja) * 2014-04-09 2017-04-13 バイオ−ラッド・イノベーションズＢｉｏ−Ｒａｄ Ｉｎｎｏｖａｔｉｏｎｓ 分析方法でシグナル検出を改善する吸収性粒子の使用

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EP0169434A2 (en) 1986-01-29
EP0169434B1 (en) 1991-09-25
US4731337A (en) 1988-03-15
DE3584196D1 (de) 1991-10-31
EP0169434A3 (en) 1987-12-02
JP2584611B2 (ja) 1997-02-26
FI842992A0 (fi) 1984-07-26
ATE67858T1 (de) 1991-10-15

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