JPH0331227B2 - - Google Patents
Info
- Publication number
- JPH0331227B2 JPH0331227B2 JP57202166A JP20216682A JPH0331227B2 JP H0331227 B2 JPH0331227 B2 JP H0331227B2 JP 57202166 A JP57202166 A JP 57202166A JP 20216682 A JP20216682 A JP 20216682A JP H0331227 B2 JPH0331227 B2 JP H0331227B2
- Authority
- JP
- Japan
- Prior art keywords
- antibody
- antigen
- reaction
- latex
- agglutination
- 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.)
- Expired - Lifetime
Links
- 239000000427 antigen Substances 0.000 claims description 25
- 102000036639 antigens Human genes 0.000 claims description 25
- 108091007433 antigens Proteins 0.000 claims description 25
- 238000006243 chemical reaction Methods 0.000 claims description 19
- 239000002245 particle Substances 0.000 claims description 16
- 230000004520 agglutination Effects 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 13
- 239000011541 reaction mixture Substances 0.000 claims description 2
- 239000004816 latex Substances 0.000 description 16
- 229920000126 latex Polymers 0.000 description 16
- 210000002966 serum Anatomy 0.000 description 15
- 229940027941 immunoglobulin g Drugs 0.000 description 8
- 238000005259 measurement Methods 0.000 description 8
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 238000004220 aggregation Methods 0.000 description 6
- 230000002776 aggregation Effects 0.000 description 6
- 241000283973 Oryctolagus cuniculus Species 0.000 description 5
- 238000010790 dilution Methods 0.000 description 5
- 239000012895 dilution Substances 0.000 description 5
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- 239000008363 phosphate buffer Substances 0.000 description 4
- 108060003951 Immunoglobulin Proteins 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 102000018358 immunoglobulin Human genes 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 230000001235 sensitizing effect Effects 0.000 description 3
- 230000000405 serological effect Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 239000004471 Glycine Substances 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 229940072221 immunoglobulins Drugs 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 101710081722 Antitrypsin Proteins 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 108010075016 Ceruloplasmin Proteins 0.000 description 1
- 102100023321 Ceruloplasmin Human genes 0.000 description 1
- 241001478240 Coccus Species 0.000 description 1
- -1 IgG Proteins 0.000 description 1
- 241000283977 Oryctolagus Species 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 206010070834 Sensitisation Diseases 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- 241000295644 Staphylococcaceae Species 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- 102000004338 Transferrin Human genes 0.000 description 1
- 108090000901 Transferrin Proteins 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000001475 anti-trypsic effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000009918 complex formation Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 238000007720 emulsion polymerization reaction Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000010419 fine particle Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 229910052809 inorganic oxide Inorganic materials 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229920002285 poly(styrene-co-acrylonitrile) Polymers 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 230000008313 sensitization Effects 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 230000007928 solubilization Effects 0.000 description 1
- 238000005063 solubilization Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000012581 transferrin Substances 0.000 description 1
- 239000002753 trypsin inhibitor Substances 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
- G01N33/53—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
Landscapes
- 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)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (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)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Description
本発明は不溶性担体粒子を用いる凝集反応測定
方法に関する。
近年、臨床検査の分野においては、ある種の疾
患を血清学的に診断することが重要視されてい
る。そして、この診断のためには、抗原及び抗体
を正確、簡便かつ迅速に定量することが、極めて
重要な課題となつている。本発明はかかる課題の
解決を目的とするものである。
一般的に、抗原−抗体はそれぞれ一定濃度の範
囲内で反応が起こり、その複合体を形成する。こ
の濃度比が極端に偏ると抗原−抗体複合体が形成
されにくくなる。一定量の抗体存在下で抗原量を
増加していくと、ある一定の抗原量まで抗原濃度
に比例して複合体形成による沈澱物は増加してい
くが、抗原濃度がある一点を越えると逆に沈降物
の量は減少していく。この減少は抗原過剰による
複合体の可溶化現象としてよく知られている。
このことは、血清学的ラテツクス試薬を用いた
抗原の測定反応においても起こりうる。血清学的
ラテツクス試薬は、通常粒径0.005〜1μのラテツ
クス粒子に抗体又は抗原を感作させ、弱アルカリ
性の緩衝液に分散させたものである。しかし、一
定量のラテツクス粒子に感作できる抗体又は抗原
量は自と限界があるため測定できる抗原又は抗体
量にも限界がある。従つて、高濃度抗原の測定に
際しては前記の抗原過剰現象の発現によつて、そ
のままでは低値に誤つて測定されることがある。
この場合、検体(サンプル)の希釈操作でその誤
りを回避しようとするが、これは煩雑である。例
えば、免疫グロブリG(IgG)の測定では検体血
清を約1000〜10000倍希釈するのが通例であり、
これは臨床検査薬の測定操作の簡略化というニー
ズを考慮すると大きな欠点となる。
従来、ラテツクスのような不溶性の担体粒子に
抗体又は抗原を感作した試薬を用いた抗原−抗体
凝集反応を行ない、その反応の結果生じた複合体
の濁度を光学的に測定する方法は提案されている
〔特開昭54−108693、特開昭55−159157、特開昭
57−1970及び沢井・石田、検査と技術:10(6)、
555(1982)など〕。しかし、これらにみられる方
法においても、検体が高濃度の場合では例外なく
大幅な希釈操作を必要とするため、希釈による大
きな誤差、測定範囲が狭いことに起因する抗原過
剰現象による誤つた測定値を出す危険もはらんで
いる。
本発明者は上記につき検討した結果、免疫グロ
ブリンのような検体血清中に存在する高濃度抗原
を測定する場合、検体中の抗原に特異的に反応し
得るフリーの抗体を不溶性担体粒子に感作した抗
体とともに反応系へ添加することにより、測定範
囲を拡大して抗原過剰現象の発現頻度を避け、以
つて検体血清の希釈操作も極力少なくし、正確で
簡単な高濃度抗原を測定する方法を見出し、本発
明を完成するに至つた。
即ち、本発明は不溶性担体粒子に抗体を感作さ
せ、これと検体とを反応させて、この反応混合物
の凝集を光学的に測定する方法において、不溶性
担体粒子に感作した抗体とは別に、検体中の抗原
に特異的に反応し得るフリーの抗体を反応系に加
えて抗原−抗体反応を行うことを特徴とする凝集
反応測定方法である。
本発明の方法によれば、凝集反応は不溶性担
体粒子を感作した抗体検体血清中の抗原と反応
し得るフリーのモノスペシフイクな抗体抗体作
製動物の正常血清検体血清とを順次、液体媒体
中で数分間振とう混合によつて反応させ、希釈液
で一定容量に希釈後、抗原−抗体反応に基づく凝
集に光を照射し、その際の吸光度の変化により定
量的に測定できる。もちろん、上記のピペツト操
作の順序は任意であり、上記番号順に拘る必要は
ないが、検体血清だけは最後に反応させた方が望
ましい。
本発明で使用する不溶性担体粒子は、有機高分
子物質の微粒子のもの、例えばポリスチレン、カ
ルボキシ変性、スルフオン酸変性、スチレンアク
リロニトリル共重合体の如き乳化重合により得ら
れるラテツクス、個々に分散されたブドウ球菌、
連鎖球菌の如き球菌型の細菌又はシリカ、シリカ
−アルミナ、アルミナの如き無機酸化物、その他
鉱物紛末、金属等が用いられる。そして、このよ
うな不溶性担体粒子に抗体を感作させる方法は公
知の方法が多くあり、これに従つて良い。また、
希釈液には生理食塩水、リン酸、トリス−塩酸等
の緩衝液を用いる。
本発明の方法で測定できる対象は、血清中の
IgG、IgA、IgM等の如き免疫グロブリン、アル
ブミン、セルロプラスミン、トランスフエリン、
アンチトリプシン等が挙げられるが、特に免疫グ
ロブリンのような検体血清中に高濃度に存在する
物質の測定に有効である。
本発明法において、不溶性担体に感作した抗体
にフリーの抗体を加えると、抗体のロツトにより
非特異凝集が起こることがある。このような場合
反応系に対抗作製に使用した動物の正常血清を加
えて、この非特異凝集を回避する。
本発明によれば、測定できる抗原量が増加でき
るし、正確性も期待できる。これに伴い、検体血
清の希釈倍数を大幅に軽減でき、測定範囲を大幅
に拡大することが可能になる。
以下、本発明を実施例により、さらに説明する
が、本発明はこれにより何ら限定されるものでは
ない。
実施例 1
(1) 抗ヒトIgG抗体の調製
家兎の産生したヒトIgGモノスペシフイツク
抗体を用いる。(抗原ヒトIgG鎖を固定したセ
フアローズ4Bのカラムに通液してアフイニテ
イクロマトグラフイーによつて精製。)
(2) 抗ヒトIgG抗体感作ラテツクス試薬の調製ラ
テツクス(日本合成ゴム社製、カルボキシ変性
タイプ、粒径0.305μ)をPH8のグリシン緩衝液
に浮遊(0.5%)させ、この浮遊液1容と、リ
ン酸緩衝液で500μg/mlに調製した抗ヒトIgG
抗体溶液1容とを混合し、室温で30分保つた
後、10000×Gで20分遠心分離して、ラテツク
ス粒子で末吸着のウサギr−グロブリン分子を
除去し、次に沈降したラテツクス粒子を再び1
%BSAを含むグリシン緩衝液に0.5%になるよ
う分散させて感作ラテツクスを調製する。
(3) ラテツクス凝集反応の測定
上記抗IgGラテツクス試薬20μ、抗IgG抗体
液20μ、リン酸緩衝液で3倍希釈の正常家兎
血清20μ及び検体血清又は標準品20μを小
試験管にとり、10分間振とう混合したのち、希
釈液4mlを加え340mmにおける吸光度を日立製
作所200−10型分光光度計を用いて測定し、ブ
ランクに対する吸光度の変化(−△340)によ
り凝集活性を求める。(濃度の算出は標準品の
−△340を検量として行なう)
比較例 1
() 実施例1の(3)でのラテツクス凝集反応の測
定において、フリーの抗体による非特異凝集防
止効果について正常家兎血清を除いて測定する
代わりに、リン酸緩衝液(100mM、PH7.5)を
用いて測定する。所定の方法により凝集反応を
行なつたのち、肉眼で凝集の有無を判定した。
その結果を表1に示す。
The present invention relates to a method for measuring agglutination reactions using insoluble carrier particles. In recent years, in the field of clinical testing, serological diagnosis of certain diseases has become important. For this diagnosis, accurate, simple, and rapid quantification of antigens and antibodies has become an extremely important issue. The present invention aims to solve this problem. Generally, an antigen-antibody reaction occurs within a certain concentration range to form a complex. If this concentration ratio is extremely biased, it becomes difficult to form an antigen-antibody complex. When the amount of antigen is increased in the presence of a certain amount of antibody, the amount of precipitate due to complex formation increases in proportion to the antigen concentration up to a certain amount of antigen, but once the antigen concentration exceeds a certain point, the opposite occurs. The amount of sediment decreases. This decrease is well known as a solubilization phenomenon of the complex due to antigen excess. This can also occur in antigen measurement reactions using serological latex reagents. Serological latex reagents are usually made by sensitizing latex particles with a particle size of 0.005 to 1 μm with antibodies or antigens and dispersing them in a weakly alkaline buffer. However, since there is a limit to the amount of antibody or antigen that can be sensitized to a certain amount of latex particles, there is also a limit to the amount of antigen or antibody that can be measured. Therefore, when measuring high-concentration antigens, the above-mentioned antigen excess phenomenon may result in erroneously measured low values.
In this case, attempts are made to avoid this error by diluting the specimen (sample), but this is complicated. For example, when measuring immunoglobulin G (IgG), it is customary to dilute the sample serum approximately 1,000 to 10,000 times.
This is a major drawback when considering the need to simplify the measurement operation of clinical test drugs. Conventionally, a method has been proposed in which an antigen-antibody agglutination reaction is performed using a reagent in which insoluble carrier particles such as latex are sensitized with antibodies or antigens, and the turbidity of the complex formed as a result of the reaction is optically measured. [Unexamined Japanese Patent Publication No. 54-108693, Unexamined Japanese Patent Application No. 55-159157, Unexamined Unexamined Publication
57-1970 and Sawai and Ishida, Inspection and Technology: 10(6),
555 (1982) etc.]. However, even with these methods, when the sample concentration is high, extensive dilution is required without exception, resulting in large errors due to dilution and erroneous measurement values due to antigen excess phenomenon caused by the narrow measurement range. There is also a risk of releasing. As a result of studying the above, the present inventor found that when measuring a high concentration antigen such as immunoglobulin present in a sample serum, insoluble carrier particles are sensitized with free antibodies that can specifically react with the antigen in the sample. By adding the antibody to the reaction system together with the antibody, the measurement range is expanded and the frequency of antigen excess phenomenon is avoided, and the dilution of the sample serum is minimized, thereby creating an accurate and simple method for measuring high-concentration antigens. This finding led to the completion of the present invention. That is, the present invention provides a method in which insoluble carrier particles are sensitized with an antibody, the antibody is reacted with a sample, and the aggregation of this reaction mixture is optically measured. This is an agglutination reaction measurement method characterized by adding a free antibody capable of specifically reacting with an antigen in a specimen to a reaction system to perform an antigen-antibody reaction. According to the method of the present invention, the agglutination reaction is performed by sequentially sensitizing insoluble carrier particles with a free monospecific antibody capable of reacting with the antigen in the antibody sample serum and the normal serum sample serum of an antibody-producing animal in a liquid medium. The reaction is caused by shaking and mixing for a minute, and after dilution to a constant volume with a diluent, the agglutination based on the antigen-antibody reaction is irradiated with light, and it can be quantitatively measured by the change in absorbance at that time. Of course, the order of the pipetting operations described above is arbitrary and there is no need to adhere to the above numerical order, but it is preferable to react only the sample serum last. The insoluble carrier particles used in the present invention are fine particles of organic polymeric substances, such as polystyrene, carboxy-modified, sulfonic acid-modified, latex obtained by emulsion polymerization such as styrene-acrylonitrile copolymers, and individually dispersed staphylococci. ,
Coccus type bacteria such as streptococcus, inorganic oxides such as silica, silica-alumina, alumina, other mineral powders, metals, etc. are used. There are many known methods for sensitizing antibodies to such insoluble carrier particles, and any of these methods may be used. Also,
A buffer solution such as physiological saline, phosphoric acid, or Tris-HCl is used as the diluent. The target that can be measured by the method of the present invention is
Immunoglobulins such as IgG, IgA, IgM, etc., albumin, ceruloplasmin, transferrin,
Examples include antitrypsin, and it is particularly effective for measuring substances such as immunoglobulins that are present in high concentrations in sample serum. In the method of the present invention, when a free antibody is added to an antibody sensitized to an insoluble carrier, nonspecific aggregation may occur due to a lot of antibodies. In such cases, the normal serum of the animal used for counterproduction is added to the reaction system to avoid this nonspecific agglutination. According to the present invention, the amount of antigen that can be measured can be increased and accuracy can also be expected. Accordingly, the dilution factor of the sample serum can be significantly reduced, and the measurement range can be significantly expanded. EXAMPLES Hereinafter, the present invention will be further explained with reference to Examples, but the present invention is not limited thereto. Example 1 (1) Preparation of anti-human IgG antibody A human IgG monospecific antibody produced in a domestic rabbit is used. (The solution was passed through a Sepharose 4B column immobilized with the antigen human IgG chain and purified by Affinitei chromatography.) (2) Preparation of anti-human IgG antibody sensitization latex reagent Latex (manufactured by Nippon Gosei Rubber Co., Ltd., carboxy Denatured type, particle size 0.305μ) was suspended (0.5%) in a pH8 glycine buffer, and 1 volume of this suspension was mixed with anti-human IgG adjusted to 500μg/ml with a phosphate buffer.
Mix with 1 volume of antibody solution, keep at room temperature for 30 minutes, centrifuge at 10,000 x G for 20 minutes to remove rabbit r-globulin molecules adsorbed on the latex particles, and then remove the precipitated latex particles. 1 again
A sensitized latex is prepared by dispersing it in a glycine buffer containing 0.5% BSA. (3) Measurement of latex agglutination reaction Add 20μ of the above anti-IgG latex reagent, 20μ of anti-IgG antibody solution, 20μ of normal rabbit serum diluted 3 times with phosphate buffer, and 20μ of sample serum or standard product into a small test tube and wait for 10 minutes. After shaking and mixing, 4 ml of the diluent was added, and the absorbance at 340 mm was measured using a Hitachi Model 200-10 spectrophotometer, and the aggregation activity was determined by the change in absorbance (-Δ340) relative to the blank. (The concentration is calculated using -△340 of the standard product as a calibration.) Comparative Example 1 () In the measurement of latex agglutination reaction in (3) of Example 1, the effect of free antibody on preventing nonspecific agglutination was evaluated using normal rabbits. Instead of measuring without serum, measure using phosphate buffer (100mM, PH7.5). After carrying out an agglutination reaction according to a predetermined method, the presence or absence of agglutination was determined with the naked eye.
The results are shown in Table 1.
【表】
() ヒト血清成分による非特異凝集防止効果に
ついて、正常家兎血清を除いて測定する代わり
にリン酸緩衝液(100mM、PH7.5)を用いて測
定する。凝集の判定は(1)に準じた。その結果を
表2に示す。尚、ラテツクス試薬は抗α−フエ
トプロテイン抗体(家兎)感作したものを使用
した。調製法は実施例1の(2)に準ずる。[Table] () The non-specific agglutination prevention effect of human serum components is measured using phosphate buffer (100mM, PH7.5) instead of excluding normal rabbit serum. Aggregation was determined according to (1). The results are shown in Table 2. The latex reagent used was one sensitized with anti-α-fetoprotein antibody (rabbit). The preparation method is based on Example 1 (2).
【表】
−:肉眼で凝集を認めないもの
+:肉眼で明らかに凝集を認めるもの
[Table] −: No aggregation observed with the naked eye +: Appearances with obvious aggregation observed with the naked eye
第1図はラテツクス試薬に抗原を添加し凝集さ
せた時の吸収スペクトルを対照(抗原無添加)の
ものと比較した図である。
FIG. 1 is a diagram comparing the absorption spectrum obtained when an antigen is added to a latex reagent and aggregated with that of a control (no antigen added).
Claims (1)
体とを反応させて、この反応混合物の凝集を光学
的に測定する方法において、不溶性担体粒子に感
作した抗体とは別に、検体中の抗原に特異的に反
応し得るフリーの抗体を反応系に加えて抗原−抗
体反応を行うことを特徴とする不溶性担体粒子を
用いる凝集反応測定方法。1 In a method in which insoluble carrier particles are sensitized with an antibody, the antibody is reacted with a specimen, and the agglutination of this reaction mixture is optically measured, the antigen in the specimen is separated from the antibody sensitized to the insoluble carrier particles. 1. A method for measuring an agglutination reaction using insoluble carrier particles, characterized in that an antigen-antibody reaction is carried out by adding a free antibody capable of specifically reacting to a reaction system.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP20216682A JPS5992353A (en) | 1982-11-19 | 1982-11-19 | Measuring method of flocculating reaction using insoluble carrier particle |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP20216682A JPS5992353A (en) | 1982-11-19 | 1982-11-19 | Measuring method of flocculating reaction using insoluble carrier particle |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS5992353A JPS5992353A (en) | 1984-05-28 |
JPH0331227B2 true JPH0331227B2 (en) | 1991-05-02 |
Family
ID=16453050
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP20216682A Granted JPS5992353A (en) | 1982-11-19 | 1982-11-19 | Measuring method of flocculating reaction using insoluble carrier particle |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS5992353A (en) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH067130B2 (en) * | 1986-01-17 | 1994-01-26 | 株式会社ミドリ十字 | Aqueous solvent for agglutination test |
JPS63115061A (en) * | 1986-10-31 | 1988-05-19 | Sekisui Chem Co Ltd | Immunoassay |
DE4309393A1 (en) * | 1993-03-23 | 1994-09-29 | Boehringer Mannheim Gmbh | Reduction of the hook effect in immunoassays with particulate carrier material |
CN103293299B (en) * | 2012-02-24 | 2016-06-08 | 珀金埃尔默医学诊断产品(上海)有限公司 | Widen method and the test kit thereof of double-antibody sandwich immunodetection concentration range |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5631648A (en) * | 1979-06-19 | 1981-03-31 | Siber George | Test in which cohesion is caused by contact with body fluid containing antigen and method of preparing reagent for said test |
JPS579723A (en) * | 1980-06-20 | 1982-01-19 | Mitsubishi Chem Ind Ltd | Stabilizing agent for immunological reaction and measuring method of antigen-antibody reaction |
JPS57111446A (en) * | 1980-12-29 | 1982-07-10 | Sekisui Chem Co Ltd | Determing method of latex agglutination |
-
1982
- 1982-11-19 JP JP20216682A patent/JPS5992353A/en active Granted
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5631648A (en) * | 1979-06-19 | 1981-03-31 | Siber George | Test in which cohesion is caused by contact with body fluid containing antigen and method of preparing reagent for said test |
JPS579723A (en) * | 1980-06-20 | 1982-01-19 | Mitsubishi Chem Ind Ltd | Stabilizing agent for immunological reaction and measuring method of antigen-antibody reaction |
JPS57111446A (en) * | 1980-12-29 | 1982-07-10 | Sekisui Chem Co Ltd | Determing method of latex agglutination |
Also Published As
Publication number | Publication date |
---|---|
JPS5992353A (en) | 1984-05-28 |
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