JPH01102353A - Immune sensor - Google Patents
Immune sensorInfo
- Publication number
- JPH01102353A JPH01102353A JP62259543A JP25954387A JPH01102353A JP H01102353 A JPH01102353 A JP H01102353A JP 62259543 A JP62259543 A JP 62259543A JP 25954387 A JP25954387 A JP 25954387A JP H01102353 A JPH01102353 A JP H01102353A
- Authority
- JP
- Japan
- Prior art keywords
- protein
- metal wire
- electrode
- silver
- dialdehyde compound
- 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
- 229910052751 metal Inorganic materials 0.000 claims abstract description 15
- 239000002184 metal Substances 0.000 claims abstract description 15
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 14
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 14
- -1 dialdehyde compound Chemical class 0.000 claims abstract description 9
- 239000000203 mixture Substances 0.000 claims abstract description 7
- 238000004132 cross linking Methods 0.000 claims abstract description 6
- 239000012528 membrane Substances 0.000 claims abstract description 5
- 229940027941 immunoglobulin g Drugs 0.000 claims description 12
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims description 3
- 229910052709 silver Inorganic materials 0.000 abstract description 8
- 239000004332 silver Substances 0.000 abstract description 8
- 229910021607 Silver chloride Inorganic materials 0.000 abstract description 7
- 238000000034 method Methods 0.000 abstract description 7
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 abstract description 7
- 238000011282 treatment Methods 0.000 abstract description 5
- 239000007788 liquid Substances 0.000 abstract 2
- 238000006243 chemical reaction Methods 0.000 description 5
- 238000003018 immunoassay Methods 0.000 description 4
- 108010088751 Albumins Proteins 0.000 description 3
- 102000009027 Albumins Human genes 0.000 description 3
- 239000000427 antigen Substances 0.000 description 3
- 102000036639 antigens Human genes 0.000 description 3
- 108091007433 antigens Proteins 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 239000012188 paraffin wax Substances 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000012085 test solution Substances 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 206010018910 Haemolysis Diseases 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000024203 complement activation Effects 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- ZOMNIUBKTOKEHS-UHFFFAOYSA-L dimercury dichloride Chemical class Cl[Hg][Hg]Cl ZOMNIUBKTOKEHS-UHFFFAOYSA-L 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000008588 hemolysis Effects 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 230000006371 metabolic abnormality Effects 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 210000004885 white matter Anatomy 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
【発明の詳細な説明】 〔産業上の利用分野〕 本発明は、免疫センサに関する。更に詳しくは。[Detailed description of the invention] [Industrial application field] The present invention relates to an immunosensor. For more details.
免疫グロブリンＧの測定に有効に用いられる免疫センサ
に関する。The present invention relates to an immunosensor that is effectively used for measuring immunoglobulin G.
抗原は、それに対応する抗体と特異的に結合し、複合体
を形成する。このため、抗原−抗体反応を利用した免疫
分析法が、特に臨床化学検査でよく用いられている。The antigen specifically binds to its corresponding antibody to form a complex. For this reason, immunoassay methods that utilize antigen-antibody reactions are often used, particularly in clinical chemistry tests.
この免疫分析には、既知の抗原を用いて対応する抗体を
測定する方法あるいは補体などの活性を定量する方法な
どがある。抗原−抗体複合体形成の判定には、沈降反応
、凝集反応、溶血反応、中 −和反応あるいは補体結合
反応などが利用されており、またラジオイムアッセイ（
ＲＩＡ）、酵素免疫法（ＥＩＡ）、蛍光免疫法など、抗
原あるいは抗体を標識する方法も広く行われているが、
これらの方法はいずれも煩雑な操作を必要とする欠点が
みられる。This immunoassay includes a method of measuring the corresponding antibody using a known antigen or a method of quantifying complement activity. Precipitation reactions, agglutination reactions, hemolysis reactions, neutralization reactions, and complement fixation reactions are used to determine the formation of antigen-antibody complexes, and radioimmune assays (
Methods for labeling antigens or antibodies, such as RIA), enzyme immunoassay (EIA), and fluorescence immunoassay, are also widely used.
All of these methods have the disadvantage of requiring complicated operations.
抗体およびこれと構造上・機能上の関連をもったん白質
である免疫グロブリンの内、たん白質代謝異常の重要な
指標である免疫グロブリンＧ（ＩｇＧ）の測定の際にも
、このような解決されるべき課題が残されており、それ
の簡易な測定法の開発が強く望まれている。Among antibodies and immunoglobulin, which is a white matter protein that is structurally and functionally related to antibodies, such unresolved problems also arise when measuring immunoglobulin G (IgG), which is an important indicator of protein metabolic abnormalities. There are still issues to be solved, and the development of a simple measurement method is strongly desired.
本発明者は、かかる従来技術の欠点を解決すべく種々検
討した結果、免疫グロブリンＧを結合するたん白質であ
るプロティンＡを金属線上に特定の手段で固定させるこ
とにより、かかる課題が効果的に解決されることを見出
した。As a result of various studies aimed at solving the drawbacks of the prior art, the present inventors found that by fixing protein A, which is a protein that binds immunoglobulin G, on a metal wire using a specific method, the problem can be effectively solved. I found a solution.
従って、本発明は免疫グロブリンＧの測定に有効に用い
られる免疫センサに係り、この免疫センサは、金属線上
に被覆されたプロティンＡ−アルブミン混合物膜をジア
ルデヒド化合物で架橋処理したものからなる。Therefore, the present invention relates to an immunosensor that is effectively used for measuring immunoglobulin G, and this immunosensor is made of a protein A-albumin mixture membrane coated on a metal wire and cross-linked with a dialdehyde compound.
金属線としては、例えば銀／塩化銀、白金、銀、銅など
の金属線であって、その太さが電極を形成し得る程度の
ものが用いられる。The metal wire used is, for example, a metal wire made of silver/silver chloride, platinum, silver, copper, etc., and whose thickness is such that it can form an electrode.
これらの金属線へのプロティンＡ−アルブミン混合物膜
の被覆は、プロティンＡを約１〜１０％、アルブミンを
約１０〜２０％の濃度でそれぞれ溶解させた水溶液中に
金属線を部分的に浸漬するなどのコーティング手段を適
用し、次いでこれを約３〜５℃で約１〜２４時間放置す
ることによって行われる。To coat these metal wires with a protein A-albumin mixture film, the metal wires are partially immersed in an aqueous solution in which protein A is dissolved at a concentration of about 1 to 10% and albumin is dissolved at a concentration of about 10 to 20%. This is done by applying a coating such as, and then allowing it to stand at about 3-5°C for about 1-24 hours.
ここでアルブミンが用いられるのは、金属線との接着性
が良く、またアミノ基を有するため、次工程でジアルデ
ヒド化合物を介してプロティンＡを架橋して固定できる
こと、更にｐＨ変化に対して緩衝能を有するためである
。Albumin is used here because it has good adhesion to metal wires, and because it has an amino group, protein A can be cross-linked and fixed via a dialdehyde compound in the next step, and it also buffers against pH changes. This is because they have the ability.
次いで、この金属線上に被覆されたプロティンＡ−アル
ブミン混合物膜を固定化するため、ジアルデヒド化合物
での架橋処理が行われる。この架橋処理は、グルタルア
ルデヒドによって代表されるジアルデヒド化合物の約０
．５〜１０％水溶液中に、約３〜５℃で約１〜２４時間
浸漬することによって行われ、固定化をより強固なもの
とさせる。Next, in order to immobilize the protein A-albumin mixture film coated on the metal wire, crosslinking treatment with a dialdehyde compound is performed. This cross-linking process results in approximately 0% of the dialdehyde compound represented by glutaraldehyde.
．． This is done by immersing in a 5-10% aqueous solution at about 3-5° C. for about 1-24 hours, thereby making the immobilization stronger.
〔作用〕および〔発明の効果〕
本発明に係る免疫センサは、金属線の一部にこのような
処理膜を設け、処理膜が設けられなかった部分をパラフ
ィンなどで被覆して免疫センサ電極とし、これに銀／塩
化銀電極、飽和カロメル電極、水素電極、前記各種金属
線などを参照電極として、これらをエレクトロメーター
に接続することにより、被験液の濃度を電位差として測
定することができる。[Function] and [Effects of the Invention] The immunosensor according to the present invention is provided with such a treated film on a part of the metal wire, and the part where the treated film is not provided is coated with paraffin or the like to be used as an immunosensor electrode. By using this as a reference electrode, such as a silver/silver chloride electrode, a saturated calomel electrode, a hydrogen electrode, or the various metal wires mentioned above, and connecting these to an electrometer, the concentration of the test solution can be measured as a potential difference.
この電位差変化は、１０−４〜１０−”ｇ／ｍ Ｑの免
疫グロブリンＧ濃度範囲で直線的な関係を示すので。This change in potential difference shows a linear relationship in the immunoglobulin G concentration range of 10-4 to 10-''g/mQ.
免疫グロブリンＧの計測に使用し得る免疫センサとして
有効に作動する。しかも、その製作は容易であり、その
操作も簡便に行ない得るという利点を備えている。It effectively operates as an immunosensor that can be used to measure immunoglobulin G. Moreover, it has the advantage of being easy to manufacture and easy to operate.
次に、実施例について本発明を説明する。 Next, the present invention will be explained with reference to examples.
実施例
長さ５ｃｍの線状銀／塩化銀電極の一部（約１ｃＩＩの
部分）を、プロティンＡ（シグマ社製品）を錦、牛血清
アルブミン（シグマ社製品）を１５％の濃度でそれぞれ
溶解させた水溶液中に浸漬し、引き上げてから４℃で８
時間放置した。次いで、濃度１％のグルタルアルデヒド
水溶液中に、４℃で８時間浸漬し、水洗した。Example: A portion of a linear silver/silver chloride electrode with a length of 5 cm (approximately 1 cII portion) was dissolved in protein A (product of Sigma) and bovine serum albumin (product of Sigma) at a concentration of 15%. 8℃ at 4℃ after being immersed in an aqueous solution.
I left it for a while. Next, it was immersed in an aqueous glutaraldehyde solution with a concentration of 1% at 4° C. for 8 hours, and washed with water.
このような一連の処理がなされなかった銀／塩化銀電極
の露出部分をパラフィンで覆って絶縁したものを免疫セ
ンサ電極として用い、銀／塩化銀電極を参照電極として
用いて、免疫グロブリンＧ濃度１０−’ 〜１０−”ｇ
／ｍＱの被験液（緩衝液としては１゜鱈のトリス−ＨＣ
Ｑを使用、ｐＨは９．０であり、免疫グロブリンＧは負
に荷電している状態となっている）について、エレクト
ロメーターにより電位差を測定した。The exposed part of the silver/silver chloride electrode that had not been subjected to this series of treatments was covered with paraffin to insulate it and used as an immunosensor electrode.The silver/silver chloride electrode was used as a reference electrode and the immunoglobulin G concentration was 10. −' ~10−”g
/mQ test solution (1° cod Tris-HC as buffer solution)
The potential difference was measured using an electrometer (pH was 9.0, and immunoglobulin G was negatively charged).
その結果は、第１図のグラフに示されるように、１Ｏ−
４〜１０−”ｇｅｍ Ｑの免疫グロブリンＧ濃度゛範囲
内では、電位差と濃度の対数値との間には直線関係が見
出された。なお、濃度１０−’ｇ／ｍ ＱでのＳ／Ｎ比
は２である。The results are as shown in the graph of FIG.
A linear relationship was found between the potential difference and the logarithm of the concentration in the immunoglobulin G concentration range of 4 to 10-'g/m Q. The N ratio is 2.
比較例
実施例において、アルブミンを用いないと電位差に応答
がみられなかった。Comparative Example In the example, no response to potential difference was observed unless albumin was used.
第１図は、実施例の免疫センサにおける免疫グロブリン
Ｇの濃度と電位差との関係を示すグラフである。FIG. 1 is a graph showing the relationship between the concentration of immunoglobulin G and the potential difference in the immunosensor of the example.
Claims (1)
合物膜をジアルデヒド化合物で架橋処理してなる免疫セ
ンサ。 ２、ジアルデヒド化合物がグルタルアルデヒドである特
許請求の範囲第１項記載の免疫センサ。 ３、免疫グロブリンＧの測定に用いられる特許請求の範
囲第１項記載の免疫センサ。[Scope of Claims] 1. An immunosensor obtained by crosslinking a protein A-albumin mixture membrane coated on a metal wire with a dialdehyde compound. 2. The immunosensor according to claim 1, wherein the dialdehyde compound is glutaraldehyde. 3. The immunosensor according to claim 1, which is used for measuring immunoglobulin G.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62259543A JPH01102353A (en) | 1987-10-16 | 1987-10-16 | Immune sensor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62259543A JPH01102353A (en) | 1987-10-16 | 1987-10-16 | Immune sensor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH01102353A true JPH01102353A (en) | 1989-04-20 |
Family
ID=17335568
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP62259543A Pending JPH01102353A (en) | 1987-10-16 | 1987-10-16 | Immune sensor |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH01102353A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2014524567A (en) * | 2011-08-05 | 2014-09-22 | ジャンセン ダイアグノスティックス，エルエルシー | A new method for molecular bonding to metal / metal oxide surfaces. |
-
1987
- 1987-10-16 JP JP62259543A patent/JPH01102353A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2014524567A (en) * | 2011-08-05 | 2014-09-22 | ジャンセン ダイアグノスティックス，エルエルシー | A new method for molecular bonding to metal / metal oxide surfaces. |
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