JP2691575B2 - How to measure immune response - Google Patents
How to measure immune responseInfo
- Publication number
- JP2691575B2 JP2691575B2 JP63212023A JP21202388A JP2691575B2 JP 2691575 B2 JP2691575 B2 JP 2691575B2 JP 63212023 A JP63212023 A JP 63212023A JP 21202388 A JP21202388 A JP 21202388A JP 2691575 B2 JP2691575 B2 JP 2691575B2
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- Prior art keywords
- polybasic acid
- salt
- reagent
- acid
- sample
- Prior art date
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- Peptides Or Proteins (AREA)
Description
【発明の詳細な説明】 〔産業上の利用分野〕 本発明は免疫反応の測定方法に関し、更に詳細には分
析装置の流路切り換え弁等の試薬、検体等と接触する部
分が主としてアルミナ等の親水性材料で構成されたもの
を用い、免疫学的方法により抗原、抗体等を測定する免
疫反応の測定方法に関する。TECHNICAL FIELD The present invention relates to a method for measuring an immune reaction, and more specifically, a portion such as a flow path switching valve of an analyzer, which is in contact with a reagent, a sample, or the like, is mainly made of alumina or the like. The present invention relates to a method for measuring an immune reaction in which an antigen, an antibody or the like is measured by an immunological method using a material composed of a hydrophilic material.
血清や尿中の種々の因子、蛋白質、ホルモン、薬物な
どの測定が種々の疾患の診断治療法の決定などに重要な
意義を持つようになつてきている。The measurement of various factors, proteins, hormones, drugs, etc. in serum and urine has come to have an important significance in determining diagnostic treatments for various diseases.
これらの特定の物質の測定法としては、物理化学的方
法、生化学的方法、免疫学的方法などがあるが、この中
でも免疫学的方法は特異性、感度に優れていることから
蛋白質、ホルモン等の測定に広く用いられつつある。Methods for measuring these specific substances include physicochemical methods, biochemical methods, immunological methods, etc. Among them, immunological methods are excellent in specificity and sensitivity, and therefore, proteins and hormones are used. It is being widely used for measurement of etc.
免疫学的測定法には、ゲル内免疫拡散法、免疫比濁
法、免疫比ろう法、酵素免疫測定法、ケイ光免疫測定
法、放射性免疫測定法など種々の方法があるが、近年、
臨床化学分析の分野における、一般に生化学自動分析装
置と呼ばれる自動分析装置の普及に伴い、前述の免疫学
的測定法の中でも免疫比濁法が、これら分析装置への適
応の容易さから注目され、この方法により種々の物質の
測定が試みられてきている。Immunological assay methods include various methods such as in-gel immunodiffusion method, immunoturbidimetric method, immunoparasitic method, enzyme immunoassay method, fluorescent immunoassay method, and radioimmunoassay method.
In the field of clinical chemistry analysis, with the spread of automatic analyzers generally called biochemical automatic analyzers, among the above-mentioned immunological assay methods, the immunoturbidimetric method has attracted attention due to its easy adaptability to these analyzers. The measurement of various substances has been attempted by this method.
免疫比濁法に用いる試薬は、抗血清又はその精製物の
適量を適当な緩衝液中、例えばリン酸緩衝液、トリス緩
衝液、グツド緩衝液などに溶解したものと、反応促進剤
としてポリエチレングリコール等を適当な緩衝液中に溶
解したものとを組み合わせるか又は、抗血清又はその精
製物と反応促進剤を適当な緩衝液中に溶解することによ
つて作られる。Reagents used in the immunoturbidimetric method are those prepared by dissolving an appropriate amount of antiserum or a purified product thereof in a suitable buffer solution such as phosphate buffer solution, Tris buffer solution, good buffer solution, and polyethylene glycol as a reaction accelerator. And the like dissolved in a suitable buffer, or by dissolving the antiserum or its purified product and the reaction accelerator in a suitable buffer.
また、これらの免疫比濁法用試薬を適用する自動分析
装置としては、種々のタイプの装置が市販されている
が、その中でも、特公昭59-22905に記載されているタイ
プの自動分析装置は、各試薬に対し独立した分注装置を
有することから、多数検体−多数項目の測定に適した自
動分析装置とされている。In addition, various types of instruments are commercially available as automatic analyzers to which these reagents for immunoturbidimetric method are applied. Among them, the automatic analyzer of the type described in Japanese Examined Patent Publication No. 59-22905 is Since it has an independent pipetting device for each reagent, it is an automatic analysis device suitable for measurement of a large number of specimens and a large number of items.
しかしながら、このタイプの分析装置においては、分
注装置の流路切り換え弁にアルミナを主成分とする親水
性材料を用いていること、又、検体の測定の依頼の無い
場合、この流路切り換え弁中に試薬が留まつていること
に起因し、前述の抗血清及びその精製物又は抗体標識物
を試薬中に含む免疫比濁法試薬、ラテツクス面積凝集比
濁法を試薬などを適用した場合、非特異的な凝集・沈澱
を生じるという現象が起こることが判明し、従つて測定
の基準となる試薬ブランク値が変動するため測定に重大
な影響を及ぼすという問題を有していた。However, in this type of analyzer, a hydrophilic material containing alumina as the main component is used for the flow path switching valve of the pipetting device, and this flow path switching valve is used unless a sample measurement is requested. Due to the fact that the reagents are retained in the reagent, the immunoturbidimetric method reagent containing the antiserum and its purified product or the antibody-labeled product described above, when applying the reagent such as the latex area agglutination turbidimetric method, It has been found that a phenomenon of non-specific aggregation / precipitation occurs, and accordingly, there is a problem that the reagent blank value, which serves as a reference for measurement, fluctuates, which seriously affects the measurement.
従つて、流路切り換え弁等、試薬が直接接触する部分
にアルミナ等の親水性材料を用いている自動分析装置に
適用しても、非特異的な沈澱（濁り）や凝集を生じるこ
とにより測定精度に影響を及ぼさない、抗原−抗体反応
による測定法の開発が求められていた。Therefore, even if it is applied to an automatic analyzer that uses a hydrophilic material such as alumina in the part where the reagent directly contacts, such as a flow path switching valve, measurement will occur due to non-specific precipitation (turbidity) or aggregation. There has been a demand for the development of a measurement method based on the antigen-antibody reaction, which does not affect the accuracy.
本発明者らは、免疫測定法における上記の問題を解決
するために種々の検討を行つた結果、非特異反応は抗血
清の種類、抗血清の対象抗原の種類に無関係に生じ、正
常動物血清でも生じることが判明した。更に、非特異沈
澱物の組成の分析を行つた結果、非特異沈澱物は免疫グ
ロブリンであり、抗血清をそのまま用いても、種々の方
法により免疫グロブリンを精製又は加工しても本現象に
対し効果がなかつたことから、本現象はアルミナなどか
らなるセラミツク等の親水性材料の表面に免疫グロブリ
ン分子が吸着、濃縮されることにより非特異的な凝集・
沈澱が生じるものであると判断された。As a result of various investigations to solve the above problems in the immunoassay, the present inventors have found that nonspecific reactions occur regardless of the type of antiserum and the type of target antigen of the antiserum, and normal animal serum But it turned out to occur. Furthermore, as a result of the analysis of the composition of the non-specific precipitate, the non-specific precipitate is immunoglobulin, and even if the antiserum is used as it is or the immunoglobulin is purified or processed by various methods, Since there is no effect, this phenomenon is caused by nonspecific aggregation / immobilization of immunoglobulin molecules by adsorption and concentration on the surface of hydrophilic material such as ceramics.
It was judged that precipitation occurred.
非特異沈澱物が免疫グロブリン、すなわち、抗体であ
ることから、免疫グロブリン類を含む溶液中の免疫グロ
ブリン類濃度を低くすることにより、見かけ上は非特異
沈澱物又は非特異凝集物の量は減少するが、当然のこと
ながらこれは本質的な問題を解決する手段とはならなか
つた。Since the non-specific precipitate is an immunoglobulin, that is, an antibody, the amount of non-specific precipitate or non-specific aggregate is apparently reduced by lowering the concentration of immunoglobulins in the solution containing immunoglobulins. But, of course, this wasn't a way to solve the underlying problem.
そこで本発明者らは、アルミナなどからなるセラミツ
ク等の親水性材料の表面に抗体分子が吸着することを防
ぐことにより問題が解決できると考え、免疫グロブリン
類を含む溶液の組成と本現象との関係につき詳細な検討
を行つた結果、免疫グロブリン類を含む溶液の塩濃度を
ただ単に増加させたり界面活性剤等を添加することでは
免疫グロブリン類の吸着を防ぐことはできないが、試薬
や検体中にアミノ基を有しない多塩基酸を配合すれば免
疫グロブリン類の、アルミナなどからなるセラミツク等
親水性材料表面への吸着を防ぐことができ、従つて非特
異的な凝集・沈澱を抑えることができるとの知見を得、
本発明を完成するに至つた。Therefore, the present inventors believe that the problem can be solved by preventing the adsorption of antibody molecules on the surface of a hydrophilic material such as ceramics made of alumina, and the composition of a solution containing immunoglobulins and this phenomenon As a result of conducting a detailed study on the relationship, it is not possible to prevent the adsorption of immunoglobulins by simply increasing the salt concentration of the solution containing immunoglobulins or by adding a surfactant, etc. If a polybasic acid having no amino group is added to the immunoglobulin, it is possible to prevent immunoglobulins from being adsorbed on the surface of a hydrophilic material such as ceramics made of alumina and the like, and thus prevent nonspecific aggregation / precipitation. I got the knowledge that I can do it,
The present invention has been completed.
すなわち、本発明は試薬又は／及び検体と直接接触す
る部分が親水性材料で構成されている分析装置を使用
し、抗原−抗体反応により検体中の目的物質を検出する
免疫学的測定法において、免疫グロブリン類を含む試薬
又は検体にアミノ基を有しない多塩基酸を添加し、該親
水性材料表面への免疫グロブリン類の吸着を抑制して非
特異的な凝集・沈澱を防止することを特徴とする免疫反
応の測定方法を提供するものである。That is, the present invention uses an analyzer in which a portion which is in direct contact with a reagent or / and a specimen is composed of a hydrophilic material, and in an immunological assay method for detecting a target substance in a specimen by an antigen-antibody reaction, Characterized by adding a polybasic acid having no amino group to a reagent or sample containing immunoglobulins to suppress the adsorption of immunoglobulins on the surface of the hydrophilic material and prevent nonspecific aggregation / precipitation The present invention provides a method for measuring an immune reaction.
本発明方法で用いるアミノ基を有しない多塩基酸とし
ては、２塩基酸以外のものであれば特に限定されるもの
ではないが、水溶性、毒性、蛋白に対する変性作用の有
無などを考慮するとリン酸及びグリセロリン酸、ピロリ
ン酸、フイチン酸などのリン酸誘導体、ホウ酸、シユウ
酸、クエン酸、フタル酸、酒石酸、マロン酸などのジカ
ルボン酸以上の多カルボン酸及びその誘導体、ポリアネ
トールスルホン酸、デキストラン硫酸などのスルホン酸
誘導体及び硫酸誘導体やポリアニオン等が適当である。The polybasic acid having no amino group to be used in the method of the present invention is not particularly limited as long as it is other than a dibasic acid, but in consideration of water solubility, toxicity, denaturing effect on protein, etc. Acids and glycerophosphoric acid, pyrophosphoric acid, phosphoric acid derivatives such as phytic acid, boric acid, oxalic acid, citric acid, phthalic acid, tartaric acid, polycarboxylic acids of dicarboxylic acids and their derivatives such as malonic acid, polyanethole sulfonic acid, Sulfonic acid derivatives such as dextran sulfuric acid, sulfuric acid derivatives and polyanions are suitable.
この多塩基酸は、そのまま試薬又は検体に添加しても
良いが、多塩基酸の塩として、多塩基酸と塩基の組
み合せとして、多塩基酸の塩と他の塩の組み合せとし
て又は多塩基酸と他の塩の組み合せとしてのいずれか
の方法で添加することが好ましい。This polybasic acid may be added to the reagent or the sample as it is, but as a salt of the polybasic acid, a combination of the polybasic acid and a base, a combination of the salt of the polybasic acid and another salt, or the polybasic acid. It is preferable to add it by any method as a combination of the above and other salts.
上記において、塩の形成又は組み合せて配合するため
に用いる塩基としては金属やアンモニウム化合物が使用
できるが、多塩基酸と組み合わせた場合の、溶解性や血
清蛋白質等との反応性等を考慮した場合Li、Na、Ｋ、R
b、Csなどの一価の金属及びアンモニウム塩が特に優れ
ている。多塩基酸と他の塩を組み合わせて使用する場合
の他の塩としては、金属やアンモニウムの塩が使用でき
るが、Li、Na、Ｋ、Rb、Csなどの一価の金属及びアンモ
ニウムとハロゲン原子の塩が好ましい。In the above, a metal or an ammonium compound can be used as a base for forming a salt or a combination thereof, but in consideration of solubility, reactivity with serum proteins and the like when combined with a polybasic acid. Li, Na, K, R
Monovalent metals such as b and Cs and ammonium salts are particularly excellent. As the other salt when the polybasic acid is used in combination with another salt, a metal or ammonium salt can be used, but monovalent metals such as Li, Na, K, Rb and Cs, and an ammonium and halogen atom. Are preferred.
本発明における多塩基酸及び／又は他の塩の配合の効
果は、使用する免疫ブロブリン類の濃度、使用する多塩
基酸と他の塩の種類及び組み合わせを選択すれば比較的
低濃度の添加でも十分に得ることができる。したがつ
て、添加する多塩基酸及び／又は他の塩の濃度を一概に
規定することは不可能であるが、例えば多塩基酸の
塩、多塩基酸と塩基を１種又は２種以上で用いる場
合、一般的には酸基のモル濃度として0.1M以上で十分な
効果が得られ、より好ましくは0.2M以上である。多塩
基酸の塩と他の塩又は多塩基酸と他の塩を組み合わせ
使用する場合、多塩基酸の塩又は多塩基酸を低濃度で使
用する場合には他の塩を高濃度で使用すれば良く、多塩
基酸の塩又は多塩基酸を高濃度で使用する場合には他の
塩を低濃度で使用すれば良い。一般的には、前者の場
合、例えば多塩基酸の塩又は多塩基酸を10mM以上、他の
塩を0.2M以上、後者の場合例えば多塩基酸の塩又は多塩
基酸を100mM以上、他の塩を50mMで使用すれば良く、好
ましくは何れの場合でも多塩基酸の塩又は多塩基酸を50
mM以上、他の塩を0.2M以上用いれば良い。フイチン酸な
どに代表される様な一分子中に多数の酸基を含む化合物
では効果が大きく通常の酸に比較し酸基の濃度としても
より低濃度で十分な効果が得られる。The effect of blending the polybasic acid and / or other salt in the present invention is that even if a relatively low concentration is added, the concentration of the immune blobulins to be used, the type and combination of the polybasic acid and the other salt to be used are selected. You can get enough. Therefore, it is impossible to unconditionally specify the concentration of the polybasic acid and / or other salt to be added, but for example, the salt of the polybasic acid, the polybasic acid and the base may be used alone or in combination of two or more. When used, a sufficient effect is generally obtained when the acid group has a molar concentration of 0.1 M or more, and more preferably 0.2 M or more. When using a salt of a polybasic acid with another salt or a combination of a polybasic acid with another salt, when using a salt of polybasic acid or a low concentration of polybasic acid, use another salt at a high concentration. It suffices to use a salt of a polybasic acid or another salt at a low concentration when the polybasic acid is used at a high concentration. Generally, in the former case, for example, a polybasic acid salt or polybasic acid is 10 mM or more, other salts are 0.2 M or more, and in the latter case, for example, a polybasic acid salt or polybasic acid is 100 mM or more, other The salt may be used at 50 mM, and in any case, the salt of the polybasic acid or the polybasic acid is preferably 50 mM.
mM or more and another salt of 0.2 M or more may be used. A compound containing a large number of acid groups in one molecule, such as phytic acid, has a great effect, and a sufficient effect can be obtained at a lower acid group concentration than that of a normal acid.
本発明における多塩基酸及び他の塩の濃度の上限につ
いても、使用する免疫グロブリン類の種類、濃度にも存
在することから、一概に規定することは出来ないが、一
般的には多塩基酸又はその塩と塩基又は他の塩とを共存
させた時の溶解性を考慮し、塩析効果などの生じない濃
度及び濃度の組み合わせで使用すれば良い。The upper limit of the concentration of the polybasic acid and other salts in the present invention is also present in the type and concentration of immunoglobulins used, so it cannot be unconditionally specified, but generally the polybasic acid is used. Alternatively, in consideration of solubility when the salt thereof and a base or other salt are coexistent, it may be used in a concentration and a combination of concentrations that do not cause salting-out effect.
例えば抗血清をそのまま用い多塩基酸の塩として例え
ば、リン酸ナトリウムを、例えばpH7.0で用いる場合に
は、リン酸ナトリウムとしては0.9M以下、より好ましく
は0.8M以下で使用すれば良い。例えば、免疫グロブリン
類として抗血清を用い、例えば、多塩基酸の塩としてリ
ン酸ナトリウム、他の塩として塩化ナトリウムを用いた
場合、リン酸ナトリウムを0.1M以下で用いる場合には、
塩化ナトリウムとしては4M程度まで用いることが出来
る。リン酸ナトリウムを0.25Mとした場合には塩化ナト
リウムは3M以下が望ましい。また、リン酸ナトリウムを
0.5Mで用いる場合には、塩化ナトリウムとしては2M以下
が望ましい。また、例えば、免疫グロブリン類として抗
血清または硫安分画法によるガンマーグロブリン分画を
用い、例えば、多塩基酸の塩としてデキストラン硫酸ナ
トリウム（分子量15000）を用いた場合には、デキスト
ラン硫酸ナトリウムとしては22mMできれば20mM以下で使
用することが望ましい。For example, when antiserum is used as it is and sodium phosphate is used as a salt of polybasic acid, for example, at pH 7.0, sodium phosphate may be used at 0.9 M or less, more preferably 0.8 M or less. For example, using antiserum as immunoglobulins, for example, sodium phosphate as a salt of polybasic acid, when using sodium chloride as another salt, when using sodium phosphate at 0.1M or less,
Up to about 4M can be used as sodium chloride. When sodium phosphate is 0.25M, sodium chloride is preferably 3M or less. In addition, sodium phosphate
When used at 0.5M, sodium chloride is preferably 2M or less. Further, for example, when antiserum or gamma globulin fraction by ammonium sulfate fractionation method is used as immunoglobulins, and, for example, when dextran sulfate sodium (molecular weight 15000) is used as the salt of polybasic acid, dextran sulfate sodium is If it is 22 mM, it is desirable to use it at 20 mM or less.
本発明方法においては、上記の如く、多塩基酸単独の
みならずその塩又はこれと塩基若しくは他の塩と組み合
せて使用することもできる。これらの場合試薬又は検体
中のpHは、使用する多塩基酸の電離定数により異なる
が、多塩基酸が第一電離でき、少なくとも第二電離が部
分的にでも起こるpH以上であることが好ましい。In the method of the present invention, as described above, not only the polybasic acid alone but also a salt thereof or a combination thereof with a base or another salt can be used. In these cases, the pH in the reagent or the sample varies depending on the ionization constant of the polybasic acid used, but it is preferable that the pH is equal to or higher than the pH at which the polybasic acid can first ionize and at least the second ionization partially occurs.
本発明の免疫反応の測定方法としては免疫非濁法やラ
テツクスなどの不溶性担体に抗体を結合させて用いるラ
テツクス免疫比濁法などが代表的であり、免疫グロブリ
ンを含む試薬又は検体に多塩基酸を配合する以外は公知
の方法に従つて実施することができる。As a method for measuring an immune reaction of the present invention, a non-immunoturbidity method or a latex immunoturbidimetric method in which an antibody is bound to an insoluble carrier such as latex is typical, and a reagent or sample containing immunoglobulin is polybasic acid. It can be carried out according to a known method except for blending.
すなわち、本発明の方法は、試薬又は検体中の抗血清
及び抗体の種類、抗体の対象抗原の種類に関係無く、免
疫グロブリンをそのまま用いた場合でも、免疫グロブリ
ンを種々の方法により精製又は加工して用いた場合で
も、免疫グロブリンを化学的又は物理的な方法で他の物
質と結合させて用いた場合でも適用することが可能であ
り、十分な効果を得ることができる。That is, the method of the present invention, regardless of the type of antiserum and antibody in the reagent or sample, the type of target antigen of the antibody, even when immunoglobulin is used as it is, the immunoglobulin is purified or processed by various methods. The present invention can be applied to both the case where the immunoglobulin is used and the case where the immunoglobulin is bound to another substance by a chemical or physical method and used, and a sufficient effect can be obtained.
本発明方法により、非特異反応が抑制される理由は、
アルミナなどのセラミツク親水性材料の表面において多
塩基酸がマトリツクスを形成し、免疫グロブリンの吸着
を防げるためと解される。The reason why the non-specific reaction is suppressed by the method of the present invention is
It is understood that the polybasic acid forms a matrix on the surface of the ceramic hydrophilic material such as alumina and prevents the adsorption of immunoglobulin.
したがつて、流路切り換え弁等試薬又は検体が直接接
触する部分にアルミナ等の親水性材料を用いている分析
装置を使用し、免疫比濁法やラテツクス免疫比濁法の試
薬を適用しても非特異的な凝集・沈澱の精製を抑えるこ
とができ、この結果、試薬ブランク値の変動を抑えるこ
とができ、各種免疫反応を精度良く測定することが可能
である。Therefore, using an analyzer that uses a hydrophilic material such as alumina in the area where the reagent or sample directly contacts the flow path switching valve, etc., and apply the reagent for the immunoturbidimetric method or the latex immunoturbidimetric method. Also, nonspecific purification of aggregation / precipitation can be suppressed, and as a result, fluctuations in the reagent blank value can be suppressed, and various immune reactions can be accurately measured.
以下、実施例により本発明の効果をさらに詳細に説明
するが、本発明は、これら実施例によつて限定されるも
のではない。Hereinafter, the effects of the present invention will be described in more detail with reference to Examples, but the present invention is not limited to these Examples.
実施例１ 装置として流路切り換え弁等、試薬が直接接触する部
分にアルミナ等の親水性材料が使用されている分析装
置、日立736型自動分析装置を用い、また、免疫グロブ
リンを含む試薬として抗血清（抗ヒトCRP血清）又は正
常ヤギ血清（以下NGSと略す）を用い、非特異的反応の
発生を調べた。試験において本現象が明確に捕えられる
ように、検体として生理食塩水を用い、試薬と親水性材
料の接触時間を２時間として、以下に示す分析条件にて
非特異的な凝集・沈澱による吸光度の変化を測定した。
すなわち、第一試薬として非特異的な凝集によつて生じ
た沈澱にほとんど影響しない生理食塩水を200μｌ用
い、検体として生理食塩水を10μｌ添加し、さらに第二
試薬として上で調製した各試薬を100μｌ注入後、濁り
に対して本分析装置において、最も高感度な波長である
340nmでの吸光度を測定した。Example 1 As an apparatus, an analyzer in which a hydrophilic material such as alumina is used in a portion which is in direct contact with a reagent such as a flow path switching valve, a Hitachi 736 type automatic analyzer is used. Occurrence of non-specific reaction was examined using serum (anti-human CRP serum) or normal goat serum (hereinafter abbreviated as NGS). In order to clearly capture this phenomenon in the test, physiological saline was used as the sample, the contact time between the reagent and the hydrophilic material was set to 2 hours, and the absorbance due to non-specific aggregation / precipitation under the following analysis conditions. The change was measured.
That is, as the first reagent, 200 μl of physiological saline that hardly affects the precipitation caused by non-specific aggregation was added, 10 μl of physiological saline was added as a sample, and each reagent prepared above was used as the second reagent. The wavelength is the most sensitive to turbidity in this analyzer after 100 μl injection
Absorbance at 340 nm was measured.
多塩基酸又はその塩とその他の塩を用いて試験した結
果を第１表に、多塩基酸と塩基を用いて試験した結果を
第２表に、比較例を第３表にそれぞれ示す。なお、各表
中、効果は次の基準に従い示した。Table 1 shows the results of the test using the polybasic acid or its salt and other salts, Table 2 shows the results of the test using the polybasic acid and the base, and Table 3 shows the comparative examples. In each table, the effects are shown according to the following criteria.
また、抗ヒトCRP血清（10％）を用いた場合の、リン
酸ナトリウム添加による吸光度プロフイールの変化を示
す図面を第１図に示す。図中のグラフは、それぞれ第１
表の試料No.1、３、４及び第３表の試料No.125に対応す
るものである。 Further, FIG. 1 shows a drawing showing changes in the absorbance profile due to addition of sodium phosphate when anti-human CRP serum (10%) was used. The graphs in the figure are the first
It corresponds to sample Nos. 1, 3 and 4 in the table and sample No. 125 in Table 3.
（測定操作） 試薬分注のための流路をしようとする試薬で洗浄
後、同試薬を吸入する。(Measurement operation) After washing the flow path for reagent dispensing with the reagent to be used, the reagent is inhaled.
この状態で２時間放置する。 Leave in this state for 2 hours.
上記測定条件にて検体測定を行い、340nmでの吸光
度を測定した。The sample was measured under the above measurement conditions, and the absorbance at 340 nm was measured.
（結果） 多塩基酸の塩、多塩基酸又はその塩と他の塩又は多塩
基酸と、塩基を組み合わせて添加したものは、すべてほ
ぼ完全に非特異的凝集・沈澱による吸光度の上昇を抑え
た。これに対し、ただ単に塩を添加したものは、ほとん
ど効果が見られなかつた。(result) All of the salts of polybasic acid, polybasic acid or its salt and other salts or polybasic acid, and a base added in combination almost completely suppressed the increase in absorbance due to non-specific aggregation / precipitation. On the other hand, the effect of simply adding the salt showed almost no effect.
実施例２ 抗血清として抗ヒトIgG血清、抗ヒトIgA血清、ヒト抗
IgM血清、抗ヒトC3血清、抗ヒトC4血清を用い、実施例
１と同一条件にて非特異反応の発生を調べた。Example 2 Anti-human IgG serum, anti-human IgA serum, human anti-serum
Using IgM serum, anti-human C3 serum and anti-human C4 serum, the occurrence of non-specific reaction was examined under the same conditions as in Example 1.
本発明の結果を第４表に、比較方法の結果を第５表に
示す。The results of the present invention are shown in Table 4, and the results of the comparison method are shown in Table 5.
この結果から明らかなように、本発明方法は抗血清の
種類によらず、優れた効果を得ることができる。 As is clear from this result, the method of the present invention can obtain excellent effects regardless of the type of antiserum.
実施例３ 抗血清として抗ヒトIgG血清を用い、実施例１と同様
にして非特異反応の発生を調べた。本発明の結果を第６
表に、比較方法の結果を第７表に示す。Example 3 Using anti-human IgG serum as the antiserum, the occurrence of non-specific reaction was examined in the same manner as in Example 1. The sixth result of the present invention
Table 7 shows the results of the comparison method.
実施例４ 試薬のpHを変化させ、実施例１と同様にして非特異反
応の発生を調べた。pHの調製は、水酸化ナトリウムを用
いておこなつた。本発明の結果を第８表に、比較方法の
結果を第９表に示す。 Example 4 The occurrence of non-specific reaction was examined in the same manner as in Example 1 while changing the pH of the reagent. The pH was adjusted using sodium hydroxide. The results of the present invention are shown in Table 8 and the results of the comparison method are shown in Table 9.
第１図は、リン酸ナトリウムの添加による吸光度プロフ
イールの変化を示す図面である。FIG. 1 is a drawing showing changes in absorbance profile due to addition of sodium phosphate.
Claims (1)
親水性材料で構成されている分析装置を使用し、抗原−
抗体反応により検体中の目的物質を検出する免疫学的測
定法において、免疫グロブリン類を含む試薬又は検体に
アミノ基を有しない多塩基酸を添加し、該親水性材料表
面への免疫グロブリン類の吸着を抑制して非特異的な凝
集・沈殿を防止することを特徴とする免疫反応の測定方
法。1. An analyzer using an analyzer in which a portion directly contacting a reagent or / and a specimen is made of a hydrophilic material,
In an immunoassay for detecting a target substance in a sample by an antibody reaction, a polybasic acid having no amino group is added to a reagent containing immunoglobulins or a sample, and immunoglobulins on the surface of the hydrophilic material are added. A method for measuring an immune reaction, which comprises suppressing adsorption to prevent non-specific aggregation / precipitation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63212023A JP2691575B2 (en) | 1988-08-26 | 1988-08-26 | How to measure immune response |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63212023A JP2691575B2 (en) | 1988-08-26 | 1988-08-26 | How to measure immune response |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0261561A JPH0261561A (en) | 1990-03-01 |
| JP2691575B2 true JP2691575B2 (en) | 1997-12-17 |
Family
ID=16615592
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63212023A Expired - Fee Related JP2691575B2 (en) | 1988-08-26 | 1988-08-26 | How to measure immune response |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2691575B2 (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003066047A (en) * | 2001-06-14 | 2003-03-05 | Matsushita Electric Ind Co Ltd | Immune reaction measuring method and immune reaction measuring reagent used therefor |
| DE60225788T2 (en) | 2001-12-27 | 2008-07-17 | Matsushita Electric Industrial Co., Ltd., Kadoma | IMMUNO REACTION MEASUREMENT PROCEDURE FOR IMMUNE REACTION MEASUREMENT |
| WO2004053489A1 (en) * | 2002-12-10 | 2004-06-24 | Matsushita Electric Industrial Co., Ltd. | Immunoreaction measurement method |
| JP4580180B2 (en) * | 2004-02-26 | 2010-11-10 | デンカ生研株式会社 | Measurement value decrease inhibitor for immunoassay and immunoassay using the same |
| JP2007180337A (en) * | 2005-12-28 | 2007-07-12 | Nichicon Corp | Electrolytic solution for driving electrolytic capacitor |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5925184B2 (en) * | 1978-12-22 | 1984-06-15 | 天野製薬株式会社 | Method for eliminating nonspecific inhibitory effects in immunoassays |
| JPS61274261A (en) * | 1985-05-30 | 1986-12-04 | Nitto Electric Ind Co Ltd | Immunological diagnosis reagent |
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1988
- 1988-08-26 JP JP63212023A patent/JP2691575B2/en not_active Expired - Fee Related
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| Publication number | Publication date |
|---|---|
| JPH0261561A (en) | 1990-03-01 |
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