JPH0224558A - Immunoassay - Google Patents
ImmunoassayInfo
- Publication number
- JPH0224558A JPH0224558A JP17468288A JP17468288A JPH0224558A JP H0224558 A JPH0224558 A JP H0224558A JP 17468288 A JP17468288 A JP 17468288A JP 17468288 A JP17468288 A JP 17468288A JP H0224558 A JPH0224558 A JP H0224558A
- Authority
- JP
- Japan
- Prior art keywords
- antigen
- antibody
- antiidio
- measured
- calibration curve
- 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.)
- Pending
Links
- 238000003018 immunoassay Methods 0.000 title claims description 20
- 239000000427 antigen Substances 0.000 claims abstract description 35
- 102000036639 antigens Human genes 0.000 claims abstract description 35
- 108091007433 antigens Proteins 0.000 claims abstract description 35
- 239000012634 fragment Substances 0.000 claims abstract description 11
- 238000000034 method Methods 0.000 claims description 13
- 238000006243 chemical reaction Methods 0.000 abstract description 13
- 238000011088 calibration curve Methods 0.000 abstract description 12
- 239000000126 substance Substances 0.000 abstract description 10
- 230000002401 inhibitory effect Effects 0.000 abstract description 7
- 238000002360 preparation method Methods 0.000 abstract description 2
- 238000010790 dilution Methods 0.000 description 12
- 239000012895 dilution Substances 0.000 description 12
- 230000000052 comparative effect Effects 0.000 description 10
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 8
- 239000002953 phosphate buffered saline Substances 0.000 description 8
- 238000005259 measurement Methods 0.000 description 7
- 238000012360 testing method Methods 0.000 description 6
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 5
- 229940098773 bovine serum albumin Drugs 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- 239000011324 bead Substances 0.000 description 4
- 238000007865 diluting Methods 0.000 description 4
- 239000003112 inhibitor Substances 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 210000004408 hybridoma Anatomy 0.000 description 2
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 description 1
- 206010003445 Ascites Diseases 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 241000276457 Gadidae Species 0.000 description 1
- 102000030902 Galactosyltransferase Human genes 0.000 description 1
- 108060003306 Galactosyltransferase Proteins 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 239000007979 citrate buffer Substances 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 239000004816 latex Substances 0.000 description 1
- 229920000126 latex Polymers 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
Description
【発明の詳細な説明】
［産業上の利用分野］
この発明は、測定精度が改善されたイムノアッセイに関
する。DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to an immunoassay with improved measurement accuracy.
［従来の技術］
イムノアッセイにおいては、種々の既知濃度の測定すべ
き抗原を含む標準検体（キャリブレータ−）を用いて検
量線を作製し、検体について測定された値を検量線にあ
てはめて検体中の測定すべき抗原の量を求める。従来の
イムノアッセイにおいては、検量線の作製のために用い
られるＩｌ？ｌＩ検体は精製された抗原を既知量含んで
いる。[Prior Art] In immunoassay, a calibration curve is prepared using standard specimens (calibrators) containing various known concentrations of the antigen to be measured, and the values measured for the specimens are applied to the calibration curve to determine the concentration of the antigen in the specimen. Determine the amount of antigen to be measured. In conventional immunoassays, Il? is used to prepare a standard curve. The II specimen contains a known amount of purified antigen.
一方、イムノアッセイにおいて、測定に供される検体は
血液等の体液であり、測定すべき抗原の他に多種雑多な
種々の物質を含んでいる。これらの種々の阻害物質が抗
原抗体反応を妨害するため、検量線に基づいて測定され
た抗原の測定値は実際の値よりも低くなる。換言すると
、検量線の直線性が悪化する。大量の検体についてイム
ノアッセイを行なう場合に、検体を希釈することなく原
液のままで測定できれば労力及び時間がかからず非常に
有利であるが、上記現象は抗原抗体反応を妨害する物質
が多く存在すればするほど顕著になるので、血液等の検
体を希釈せずに原液のままで測定する場合に特に問題に
なる。On the other hand, in an immunoassay, the specimen to be measured is a body fluid such as blood, which contains a wide variety of various substances in addition to the antigen to be measured. Since these various inhibitory substances interfere with the antigen-antibody reaction, the measured value of the antigen based on the calibration curve will be lower than the actual value. In other words, the linearity of the calibration curve deteriorates. When performing immunoassays on large amounts of samples, it would be very advantageous to be able to measure the sample in its original form without diluting it, as it would save time and effort.However, the above phenomenon is caused by the presence of many substances that interfere with the antigen-antibody reaction. This becomes more noticeable as the temperature increases, so it becomes a particular problem when measuring a sample such as blood without diluting it as an undiluted solution.
この問題は、もし、検量線の作製に用いる標準検体とし
て、検体と同様な種々の阻害物質を含む系の液を用いて
検量線を作製すれば解決できるものと考＾られるが、こ
れらの阻害物質は通常単一のものではなく、これらを同
定して目的とする濃度に調製することは極めて困難であ
る。It is thought that this problem could be solved if the calibration curve was prepared using a system solution containing various inhibitory substances similar to the sample as the standard sample used for preparing the calibration curve, but these inhibitors Substances are usually not a single substance, and it is extremely difficult to identify them and prepare them to the desired concentration.
［発明が解決しようとする問題点コ
従って、この発明の目的は、検体を希釈せずにＲ腋のま
まで測定した場合にも、測定値か実際の値よりも実質的
に低くならず、正確に実際の値を測定することができる
イムノアッセイを提供することである。[Problems to be Solved by the Invention] Accordingly, an object of the present invention is to provide a method in which the measured value is not substantially lower than the actual value even when the sample is measured in the R armpit without being diluted; The objective is to provide an immunoassay that can accurately measure actual values.
［問題点を解決するための手段］
本願発明者らは、鋭意研究の結果、検量線を作製する際
に、標準検体中に、測定すべき抗原に対応する抗体に対
する抗イディオ抗体を存在させると、抗原と抗イディオ
抗体とが競合することにより抗原抗体反応が妨害され、
実際の検体と同じような系が得られ、従って、抗原の測
定値が実際の値よりも小さくなることを防止することが
できることを見出しこの発明を完成した。[Means for Solving the Problem] As a result of intensive research, the inventors of the present application have discovered that when preparing a calibration curve, an anti-idio antibody against the antibody corresponding to the antigen to be measured is present in the standard sample. , the antigen-antibody reaction is interfered with by competition between the antigen and the anti-idio antibody,
The inventors have completed this invention by discovering that a system similar to that of an actual specimen can be obtained and, therefore, it is possible to prevent the measured value of the antigen from being smaller than the actual value.
すなわち、この発明は、イムノアッセイにおいて、測定
すべき抗原と、該抗原に対する抗体に対する抗イディオ
抗体又はその可変領域フラグメントとを含む標準検体を
用いて検量線の作製を行なうことを特徴とする方法を提
供する。That is, the present invention provides a method for preparing a standard curve in an immunoassay using a standard sample containing an antigen to be measured and an anti-idio antibody or a variable region fragment thereof against an antibody against the antigen. do.
［発明の効果］
この発明のイムノアッセイによると、測定すべき抗原と
それに対応する抗体との抗原抗体反応において、抗イデ
ィオ抗体が抗原と競合するのて、抗原抗体反応が妨害さ
れ、ａ定に供される血液等の検体と同じような系がもた
らされる。従って、イムノアッセイにより測定される抗
原の測定値が実際の値よりも小さくなることが防止され
、正確な抗原量を測定することができる。特に、従来の
イムノアッセイと比較して、検体を原液のまま測定する
場合に測定精度が大きく向上したので、大量の検体につ
いて測定を行なう場合に検体を希釈せずに測定すること
ができ、時間及び労力を節約することができ有利である
。[Effects of the Invention] According to the immunoassay of the present invention, in the antigen-antibody reaction between the antigen to be measured and its corresponding antibody, the anti-idioantibody competes with the antigen, so the antigen-antibody reaction is hindered and a A system similar to that used for samples such as blood is provided. Therefore, the antigen value measured by immunoassay is prevented from being smaller than the actual value, and the amount of antigen can be accurately measured. In particular, compared to conventional immunoassays, the measurement accuracy has been greatly improved when measuring samples in their undiluted state, making it possible to measure large amounts of samples without diluting them, which saves time and This is advantageous because it saves labor.
［発明の詳細な説明］
この発明のイムノアッセイは、検量線を作製する際に用
いる標準検体中に抗イディオ抗体又はその可変領域フラ
グメントを存在させることを特徴とする。抗イディオ抗
体は、抗原に対応する抗体の可変領域をエピトープとす
る抗体である。[Detailed Description of the Invention] The immunoassay of the present invention is characterized in that an anti-idio antibody or a variable region fragment thereof is present in a standard sample used when preparing a standard curve. Anti-idio antibodies are antibodies whose epitope is the variable region of an antibody corresponding to an antigen.
従って、抗イディオ抗体は、抗原に対応する抗体の可変
領域と抗原抗体反応により結合する性質を有する。一方
、抗原もその対応する抗体の可変領域と抗原抗体反応に
より結合する。従って、抗原と抗イディオ抗体とは、抗
原に対応する抗体との抗原抗体反応において競合し、こ
のため、抗イディオ抗体は抗原に対して阻害物質として
働く、このようにして１種々の阻害物質か共存する現実
の検体と同様な系が得られ、実際の値と良く一致した、
直線性の優れた検量線を得ることができる。Therefore, anti-idio antibodies have the property of binding to the variable region of an antibody corresponding to an antigen through an antigen-antibody reaction. On the other hand, antigens also bind to the variable regions of their corresponding antibodies through antigen-antibody reactions. Therefore, the antigen and the anti-idio antibody compete in the antigen-antibody reaction with the antibody corresponding to the antigen, and therefore the anti-idio antibody acts as an inhibitor against the antigen, thus one of the various inhibitors A system similar to the coexisting real samples was obtained, and the actual values matched well.
A calibration curve with excellent linearity can be obtained.
この発明のイムノアッセイにおいて用いられる抗イディ
オ抗体は、モノクローナル抗体であることか好ましい。The anti-idioantibodies used in the immunoassay of this invention are preferably monoclonal antibodies.
標準検体中に抗原と共に含まれるものは抗イディオ抗体
自身てあってもよいし、Ｆｍｂフラグメント等の可変領
域のフラグメントてあってもよい
検量線を作製するための標準検体中の抗イデイオ抗体の
濃度は、特に限定されないが、通常１ ｎｇ／ｍｌない
し１００ ｐＨ／ｍｌ程度であり、好ましくは１０ ｎ
ｇ／■１ ないしＩＯ７ｚｇ ／１１程度である。また
、標準検体中に抗イディオ抗体の可変領域フラグメント
が含まれる場合には、その濃度は通常ｌ ｎｇ／ｌない
し１００ μｇ／ｌａｌであり、好ましくは１０ｎｇ／
ｍｌ ないし１０弘ｇ／■１ 程度である。The anti-idio antibody itself may be contained together with the antigen in the standard sample, or it may be a variable region fragment such as an Fmb fragment.Concentration of the anti-idio antibody in the standard sample for preparing a standard curve. is not particularly limited, but is usually about 1 ng/ml to 100 pH/ml, preferably 10 n
g/■1 to IO7zg/11. Furthermore, when the standard sample contains variable region fragments of anti-idio antibodies, the concentration is usually 1 ng/l to 100 μg/l, preferably 10 ng/l.
It is about ml to 10 hirog/■1.
この発明のイムノアッセイに用いられる抗体は従来と同
様であり、測定精度の観点からモノクローナル抗体であ
ることが好ましい。The antibodies used in the immunoassay of this invention are the same as conventional antibodies, and from the viewpoint of measurement accuracy, monoclonal antibodies are preferred.
この発明のイムノアッセイにより測定される抗原は、い
かなる抗原であってもよい。The antigen measured by the immunoassay of this invention may be any antigen.
この発明のイムノアッセイはいかなる様式のものであっ
てもよく、従って、１１１合法でもサンドイフチ法でも
よい、また、ＥＩＡ、ＲＩＡ、ＦＩＡ、ラテックス凝集
法等、いかなる種類のイムノア・シャイであってもよい
。The immunoassay of this invention may be of any format, and therefore may be of any type, such as the 111 method or the sandwich method, or any type of immunoshy method such as EIA, RIA, FIA, latex agglutination method, etc.
［実施例］
以下、この発明を実施例に基づいて詳細に説明するが、
この発明は下記実施例に限定されるものではない。[Examples] This invention will be explained in detail based on Examples below.
This invention is not limited to the following examples.
以下の実施例において、ＧＴ−ＩＩ（ガラクトース転移
酵素イソ酵素−■）を測定するためのエンザイムイムノ
アッセイにおける本発明の例について述べる。In the following example, an example of the present invention in an enzyme immunoassay for measuring GT-II (galactosyltransferase isoenzyme-■) is described.
抗イディオモノクローナル抗体ＭＡｂ ３８７２は特開
昭６２−１７４１００号に記載の方法により作製され。Anti-idiomonoclonal antibody MAb 3872 was produced by the method described in JP-A-62-174100.
これを産生するハイブリドーマがＡＴＣＣに寄託されて
おり、その受託番号はＩＩ８８９４５である。−方、Ｍ
Ａｂ ３８７２に対する抗イディオモノクローナル抗体
ＭＡり ５７１４は特願昭６２−４９１４８号記載の方
法により作製され、これを産生ずるハイブリトーマか微
工研に寄託され、その受託番号は微工研条寄第１７５９
号である。 ＭＡｂ ５７１４のＦＩｌｂフラグメント
は。A hybridoma producing this has been deposited with the ATCC, and its accession number is II88945. - Ho, M
Anti-idiomonoclonal antibody MA5714 against Ab 3872 was produced by the method described in Japanese Patent Application No. 62-49148, and the hybridoma that produced it was deposited with FIKEN, and its accession number was FEIKEN Article No. 1759.
This is the number. The FIlb fragment of MAb 5714 is.
Ｍｏｎｏｃｌｏｎａｌ Ａｎ口ｂｏｄｉｅＳ： Ｐ
ｒ１ｎｃｉｐｌｅｓ ａｎｄＰｒａｃＬｉｃｅ 、
Ａｃａｄｅｍｉｃ Ｐｒｅｓｓ、 ｌｒ＋ｒ：、第１２
０頁に記載された方法により調製された。Monoclonal An mouth bodyS: P
r1nciples andPracLice,
Academic Press, lr+r:, 12th
Prepared by the method described on page 0.
施例１．ｌ
ａ）標準検体の調製
癌患者腹水より精製されたＧＴ−ＩＩを５％ウシ血清ア
ルブミン（ＢＳＡ）及び１００ ｎｇ／ｍｌのＭＡｂ
５７１４ Ｆ ａｈフラグメントを含むリン酸緩衝食
塩水（ＰＢＳ）に、所定濃度（０，２０，５０，−１５
０ＩＩ／ｍｌ）に溶解し、標準検体とした（実施例１）
。Example 1. l a) Preparation of standard specimen GT-II purified from ascites of a cancer patient was mixed with 5% bovine serum albumin (BSA) and 100 ng/ml MAb.
5714 F ah fragment in phosphate buffered saline (PBS) at predetermined concentrations (0, 20, 50, -15
0II/ml) and used as a standard specimen (Example 1)
.
一方、比較のため、 ＭＡｂ ５７１４ Ｆ−ｂフラ
グメントを含まない５％ＢＳＡ含有ＰＢＳに同濃度のＧ
Ｔ −１１を溶解し、標準検体とした（比較例１）、
さらに、対照として、ＧＴ−■を含まない正常人プール
血清に上記濃度のＧＴ−ＩＩを溶解して標準検体とした
（対照）。On the other hand, for comparison, the same concentration of G was added to PBS containing 5% BSA without MAb 5714 F-b fragment.
T-11 was dissolved and used as a standard specimen (Comparative Example 1),
Further, as a control, GT-II at the above concentration was dissolved in normal human pooled serum not containing GT-■ and used as a standard specimen (control).
ｂ）ＧＴ−■エンザイムイムノアッセイ精製したＭＡｂ
：１８７２１０ ＪＬｇ／腸１１００■緘炭酸バツフ
アー（ｐＨ９，５）中、１７４インチのプラスチックビ
ーズを４℃、ｌ昼夜固定化した。ＰＢＳで３回洗浄後、
１％ＢＳＡ含有ＰＢＳ中、３７℃、２日間放置し、安定
化した。b) GT-■ enzyme immunoassay purified MAb
: 187210 JLg/intestine 1100 ml 174 inch plastic beads were immobilized at 4° C. day and night in carbonated buffer (pH 9,5). After washing three times with PBS,
It was stabilized by standing in PBS containing 1% BSA at 37°C for 2 days.
このビーズ１偏に上記標準検体５０＃Ｉ及び２００ ｍ
Ｍ炭酸バッフ ｙ （ｐＨ９，５） ２００ｐ Ｉを
加え、室温で１８時間放置した。３回ＰＢＳでビーズを
洗浄した後、１ｇｇ／ｍｌの１ｌｌａｂ ３８７２−Ｉ
ＩＲＰ （ホースラデイツシュペルオキシダーゼ）結合
体溶液（１％ＢＳＡ含有Ｐ Ｂ Ｓ ） ２００ルＩを
加え、室温で２時間放置した。Add the above standard sample 50 #I and 200 m to one portion of these beads.
200p I of M carbonate buffer (pH 9,5) was added, and the mixture was left to stand at room temperature for 18 hours. After washing the beads three times with PBS, 1 gg/ml 1llab 3872-I
200 lI of IRP (horseradish peroxidase) conjugate solution (PBS containing 1% BSA) was added and left at room temperature for 2 hours.
ＰＢＳて４回ビーズを洗浄後、０〜フエニレンジアミン
３ ｍｇ／ｍｌクエン酸バッファー（ｐＨ５，ｏ）５
００ｕＬｌを加え室温て３０分間発色反応させた。After washing the beads four times with PBS, 0 to phenylenediamine 3 mg/ml citrate buffer (pH 5, o) 5
00 uL was added and a color reaction was allowed to occur at room temperature for 30 minutes.
ＩＮ硫酸２鱈を加えて反応を停止させた後、４９０ｎ−
の吸光度を分光光度計により測定した。結果を表１に示
す。After stopping the reaction by adding 2 cods of IN sulfuric acid, 490n-
The absorbance of the sample was measured using a spectrophotometer. The results are shown in Table 1.
表１
以上の結果から、抗イディオ抗体を含む標準検体を用い
た本発明の実施例１の測定結果が実際の血清を用いて調
製した対照の標準検体を用いた場合の結果と非常によく
一致し・ており、一方、抗イディオ抗体を含まない標準
検体を用いた比較例１における測定結果は、実際の血清
を用いた対照の結果からかなりずれていることがわかる
。従って、本発明の方法によると、血清等の実際の検体
を測定した場合により正確な結果が得られることがわか
る。Table 1 From the above results, the measurement results of Example 1 of the present invention using the standard sample containing anti-idio antibodies are in very good agreement with the results when using the control standard sample prepared using actual serum. On the other hand, it can be seen that the measurement results in Comparative Example 1 using a standard specimen that does not contain anti-idio antibodies deviate considerably from the results of the control using actual serum. Therefore, it can be seen that according to the method of the present invention, more accurate results can be obtained when measuring an actual specimen such as serum.
実施例２、比較例２
希釈試験
阻害物質による阻害効果は、検体の希釈倍率を大きくす
ればするほど阻害物質が希釈されるので弱まり、測定結
果は真の値に近づく、逆に、希釈倍率が低い場合には、
阻害物質の影響を大きく受け、測定値は真の値よりも低
くなる。従って、検体を種々の希釈倍率で希釈して抗原
量を測定し、その結果を希釈倍率を横軸にとってプロッ
トした場合に、低希釈倍率における測定結果か、高希釈
倍率の領域の直線を外挿した直線から大きくずれている
ほど真の値と測定された値との誤差か大きいことになり
、そのアッセイの請度が低いことを示す、この例では、
これを調べるために希釈試験を行なつた。Example 2, Comparative Example 2 Dilution test The inhibitory effect of an inhibitory substance becomes weaker as the dilution rate of the specimen increases, as the inhibitory substance is diluted, and the measurement result approaches the true value. If it is low,
It is greatly influenced by inhibitors and the measured value will be lower than the true value. Therefore, when measuring the amount of antigen by diluting a sample at various dilution rates and plotting the results with the dilution rate as the horizontal axis, it is possible to extrapolate the measurement results at low dilution rates or a straight line in the region of high dilution rates. In this example, the greater the deviation from the straight line, the greater the error between the true value and the measured value, indicating that the assay is less reliable.
A dilution test was conducted to investigate this.
実施何重及び比較例１のそれぞれの機準検体を用いて検
量線を描き、希釈試験を行なフた。検体はＧ Ｔ −Ｉ
Ｔ高値の癌患者血清を用い、希釈液としてはそれぞれの
ＯＵ／Ｗｌを用いた。結果を図に示す。A calibration curve was drawn using each of the standard samples of the experimental replicates and Comparative Example 1, and a dilution test was conducted. The specimen is GT-I
Cancer patient serum with high T value was used, and each OU/Wl was used as the diluent. The results are shown in the figure.
図から明らかなように、本発明の実施例２では低希釈倍
率における測定値が、高希釈倍率における測定値を外挿
した直線上にほぼのりているのに対し、比較例２ではこ
の直線よりもかなり下にある。従ワて、この発明の方法
は比較例の方法に比べて精度が大きく優れていることが
わかる。As is clear from the figure, in Example 2 of the present invention, the measured values at low dilution ratios are almost on the straight line obtained by extrapolating the measured values at high dilution ratios, whereas in Comparative Example 2, the measured values are on the straight line extrapolated from the measured values at high dilution ratios. is also quite low. Therefore, it can be seen that the method of the present invention is significantly superior in accuracy to the method of the comparative example.
実施例３． 較例３
添加回収試験
実施例１及び比較ｌにおけるそれぞれのａｌｌ準検体を
用いて、以下の添加回収試験を行なった。Example 3. Comparative Example 3 Addition and Recovery Test The following addition and recovery test was conducted using all quasi-analytes in Example 1 and Comparison 1.
３つの検体Ａ、Ｂ又はＣ１重量部に対し、下記表２及び
３に示す濃度の標準検体１重量部を添加し、試料を作製
した。イムノアッセイ操作は実施例１と同様にして行な
った。「回収率（％）」は実測値／理論値ｘ １００
（ｚ）て与えられ、これが１００％に近いほど測定が正
確であることを示す、結果を下記表２（比較例３）及び
表３（実施例３）に示す。To 1 part by weight of the three specimens A, B, or C, 1 part by weight of a standard specimen having a concentration shown in Tables 2 and 3 below was added to prepare samples. Immunoassay operations were performed in the same manner as in Example 1. "Recovery rate (%)" is actual value/theoretical value x 100
The results are shown in Table 2 (Comparative Example 3) and Table 3 (Example 3) below.
表２及び３から、本発明の実施例３では、比較例３に比
べて回収率がはるかに１００％に近く、従って精度が高
いことがわかる。From Tables 2 and 3, it can be seen that in Example 3 of the present invention, the recovery rate was much closer to 100% than in Comparative Example 3, and therefore the accuracy was high.
図は本発明の実施例２及び比較例２についての希釈試験
の結果を示す。The figure shows the results of the dilution test for Example 2 of the present invention and Comparative Example 2.
Claims (1)
対する抗体に対する抗イディオ抗体又はその可変領域フ
ラグメントとを含む標準検体を用いて検量線の作製を行
なうことを特徴とする方法。1. A method for preparing a standard curve in an immunoassay using a standard sample containing an antigen to be measured and an anti-idio antibody or a variable region fragment thereof against an antibody against the antigen.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP17468288A JPH0224558A (en) | 1988-07-12 | 1988-07-12 | Immunoassay |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP17468288A JPH0224558A (en) | 1988-07-12 | 1988-07-12 | Immunoassay |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0224558A true JPH0224558A (en) | 1990-01-26 |
Family
ID=15982850
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP17468288A Pending JPH0224558A (en) | 1988-07-12 | 1988-07-12 | Immunoassay |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0224558A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0278177U (en) * | 1988-12-05 | 1990-06-15 |
-
1988
- 1988-07-12 JP JP17468288A patent/JPH0224558A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0278177U (en) * | 1988-12-05 | 1990-06-15 |
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