JP3411470B2 - Protein immobilization method - Google Patents
Protein immobilization methodInfo
- Publication number
- JP3411470B2 JP3411470B2 JP10206497A JP10206497A JP3411470B2 JP 3411470 B2 JP3411470 B2 JP 3411470B2 JP 10206497 A JP10206497 A JP 10206497A JP 10206497 A JP10206497 A JP 10206497A JP 3411470 B2 JP3411470 B2 JP 3411470B2
- Authority
- JP
- Japan
- Prior art keywords
- sheet
- porous carrier
- antibody
- immobilizing
- protein
- 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 - Fee Related
Links
Landscapes
- Sampling And Sample Adjustment (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、各種医療診断に利
用される免疫クロマトグラフィー装置に収納されるシー
ト状多孔質担体に、タンパク質を固定化する方法に関す
るものである。TECHNICAL FIELD The present invention relates to a method for immobilizing a protein on a sheet-shaped porous carrier housed in an immunochromatography device used for various medical diagnoses.
【０００２】[0002]
【従来の技術】各種医療機関で使用されている免疫クロ
マトグラフィー装置は、免疫学的抗原抗体反応を利用し
て、試料液中に含まれる検体の検出を行うものであり、
その結果は、医療診断に活用されている。免疫クロマト
グラフィー装置は、シート状多孔質担体とこれを収納す
るケーシングから構成される。ケーシングは、扁平な直
方体であり、その一方の端部には試料液を導入する開口
部が設けられ、もう一方の端部近くの上面には判定確認
窓が設けられている。ケーシング中に収納されているシ
ート状多孔質担体には、試料液を導入する開口部の近く
に検体を標識する抗体が保持され、判定確認窓の位置に
検体を固定化する抗体が固定されている。従来、免疫ク
ロマトグラフィー装置において、検体を固定化する抗体
をシート状多孔質担体に固定する場合、臭化シアンのよ
うな有毒試薬を用いて多段階で前処理することが必要で
あった。2. Description of the Related Art An immunochromatography device used in various medical institutions uses an immunological antigen-antibody reaction to detect a sample contained in a sample solution.
The result is used for medical diagnosis. The immunochromatography device is composed of a sheet-like porous carrier and a casing that houses it. The casing is a flat rectangular parallelepiped, and an opening for introducing the sample liquid is provided at one end of the casing, and a determination confirmation window is provided on the upper surface near the other end. The sheet-like porous carrier contained in the casing holds an antibody that labels the sample near the opening for introducing the sample solution, and an antibody that fixes the sample is fixed at the position of the determination confirmation window. There is. Conventionally, in the case of immobilizing an antibody for immobilizing a sample on a sheet-shaped porous carrier in an immunochromatography device, it was necessary to perform pretreatment in multiple steps using a toxic reagent such as cyanogen bromide.
【０００３】[0003]
【発明が解決しようとする課題】この多段階を要する前
処理には、危険な劇薬を扱ったり、洗浄を施したりしな
ければならず、安全性と操作性に大きな問題があった。
本発明は、シート状多孔質担体に、抗体などのタンパク
質を安全かつ容易に固定化する方法を提供することを目
的とする。In this pretreatment which requires multiple steps, it is necessary to handle a dangerous powerful drug and to wash it, which is a serious problem in safety and operability.
An object of the present invention is to provide a method for safely and easily immobilizing a protein such as an antibody on a sheet-shaped porous carrier.
【０００４】[0004]
【課題を解決するための手段】上記課題に鑑み、本発明
は、シート状多孔質担体上に、タンパク質溶液を滴下ま
たは転写した後、風乾または凍結乾燥させることのみか
らなるタンパク質の固定化方法であって、前記シート状
多孔質担体が、ガラス繊維、綿およびパルプからなる群
より選ばれた少なくとも１つであることを特徴とするタ
ンパク質の固定化方法に関する。これにより、タンパク
質を、安全、容易にシート状多孔質担体上に強固に固定
することができる。In view of the above problems, according to the present invention, a protein solution is dropped or transferred onto a sheet-like porous carrier and then air-dried or freeze-dried .
The method for immobilizing a protein according to claim 1, wherein the sheet-like porous carrier is at least one selected from the group consisting of glass fiber, cotton and pulp. Thus, the protein can be safely and easily firmly fixed onto the sheet-shaped porous carrier.
【０００５】[0005]
【発明の実施の形態】風乾または凍結乾燥の２つの乾燥
方法の中でも、凍結乾燥は、タンパク質の活性を低下さ
せることが少なく、特に好適に用いられる。タンパク質
は、その溶液をシート状多孔質担体にペンをもちいて塗
布（滴下）したり、フェルト等を用いてスタンプ形式で
塗布することができる。シート状多孔質担体に固定され
るタンパク質には、酵素、抗原、抗体などを用いること
ができる。検体を固定化する抗体には、マウスやラッ
ト、ウサギ、ヤギ、ヒツジ、ニワトリ、ウシなどのモノ
クロナール抗体、ポリクローナル抗体が用いられる。標
識付き抗体には、これらの抗体を金コロイド、ラテック
ス等の有色ゾルまたは水溶性色素で標識したものを用い
ることができる。このような検体を固定化する抗体およ
び標識付き抗体を用いて、尿、全血、血しょう、腹水、
唾液などの体液等を測定することができる。シート状多
孔質担体には、ガラス繊維、ポリエステル繊維、綿およ
びパルプからなる群より選ばれた少なくとも１種の不織
布あるいは抄造体が用いられる。BEST MODE FOR CARRYING OUT THE INVENTION Of the two drying methods, air drying and freeze drying, freeze drying is particularly preferable because it hardly reduces the activity of the protein. The protein can be applied (dropped) to the sheet-shaped porous carrier with a pen using a pen, or can be applied in a stamp form using a felt or the like. An enzyme, an antigen, an antibody or the like can be used as the protein immobilized on the sheet-shaped porous carrier. As an antibody for immobilizing a sample, a monoclonal antibody or a polyclonal antibody of mouse, rat, rabbit, goat, sheep, chicken, cow, etc. is used. As the labeled antibody, those labeled with a colored sol such as gold colloid or latex or a water-soluble dye can be used. By using antibodies that immobilize such samples and labeled antibodies, urine, whole blood, plasma, ascites,
Body fluids such as saliva can be measured. As the sheet-like porous carrier, at least one kind of non-woven fabric or paper product selected from the group consisting of glass fiber, polyester fiber, cotton and pulp is used.
【０００６】[0006]
【実施例】図１は、免疫クロマトグラフィー装置内部に
収納されるシート状多孔質担体の一実施例における斜視
図である。シート状多孔質担体１に、標識付き抗体２を
保持する部分、検体を固定化する抗体３を固定した部分
およびシート状多孔質担体の一部を厚くしたシンク４が
設けられている。シンク４は、流れてきた試料液を吸収
する役割がある。５は、試料液添加部である。図２は、
他の実施例におけるシート状多孔質担体の側面図であ
る。検体を固定化する抗体３を固定したシート状多孔質
担体８を、２つの基板６、標識付き抗体２を保持するシ
ート状多孔質担体７、およびシンク４を有するシート状
多孔質担体９で挟むようにして組み込んである。EXAMPLE FIG. 1 is a perspective view of an example of a sheet-like porous carrier housed inside an immunochromatography apparatus. The sheet-like porous carrier 1 is provided with a portion for holding the labeled antibody 2, a portion for fixing the antibody 3 for immobilizing the sample, and a sink 4 in which a part of the sheet-like porous carrier is thickened. The sink 4 has a role of absorbing the flowing sample liquid. Reference numeral 5 is a sample solution adding section. Figure 2
It is a side view of the sheet-like porous carrier in other examples. A sheet-shaped porous carrier 8 on which an antibody 3 for immobilizing a sample is fixed is sandwiched between two substrates 6, a sheet-shaped porous carrier 7 holding the labeled antibody 2 and a sheet-shaped porous carrier 9 having a sink 4. It is incorporated as it is.
【０００７】検体を固定化する抗体である抗ｈＣＧ抗体
の溶液にペンを浸し、この溶液をペン先からガラス繊維
濾紙に滴下した後、ガラス繊維濾紙を冷却して抗ｈＣＧ
抗体を凍結乾燥させた。さらに、抗ｈＣＧ抗体を金コロ
イドで標識した標識付き抗体を、別のガラス繊維濾紙に
保持させた。これらを図２に示すように組み込んだ。図
２における試料液添加部分５に尿をピペットで添加し
た。尿は、ガラス繊維濾紙中を浸透し、尿中に含まれて
いる抗原（ｈＣＧ）により、検体を固定化する抗体を固
定する部分３が赤紫色に変わった。このように、検体を
固定化する抗体は、ガラス繊維濾紙に、非常に強固に保
持されており、試料液によって、移動するようなことは
なかった。A pen is dipped in a solution of an anti-hCG antibody which is an antibody for immobilizing a sample, the solution is dropped from a pen tip onto a glass fiber filter paper, and then the glass fiber filter paper is cooled to cool the anti-hCG antibody.
The antibody was lyophilized. Further, the labeled antibody obtained by labeling the anti-hCG antibody with gold colloid was held on another glass fiber filter paper. These were assembled as shown in FIG. Urine was pipetted into the sample solution addition portion 5 in FIG. Urine penetrated through the glass fiber filter paper, and due to the antigen (hCG) contained in the urine, the portion 3 for fixing the antibody that immobilizes the sample turned red purple. As described above, the antibody for immobilizing the sample was held very firmly on the glass fiber filter paper, and did not move due to the sample solution.
【０００８】[0008]
【発明の効果】以上のように、本発明によれば、タンパ
ク質を安全、容易に固定化できる。またクロマトグラフ
ィー材料を選ばないことから、短時間、高精度の測定が
可能で、低コストのクロマトグラフィー装置を構成する
ことができる。INDUSTRIAL APPLICABILITY As described above, according to the present invention, proteins can be immobilized safely and easily. Further, since any chromatography material can be selected, high-accuracy measurement can be performed in a short time, and a low-cost chromatography device can be constructed.
【図１】本発明の一実施例における免疫クロマトグラフ
ィ−装置内に収納されたシート状多孔質担体の斜視図で
ある。FIG. 1 is a perspective view of a sheet-shaped porous carrier housed in an immunochromatography device according to an embodiment of the present invention.
【図２】本発明の他の実施例における免疫クロマトグラ
フィ−装置内に収納されたシート状多孔質担体の側面図
である。FIG. 2 is a side view of a sheet-shaped porous carrier housed in an immunochromatography device according to another embodiment of the present invention.
１ シート状多孔質担体
２ 標識付き抗体の保持部
３ 固定化抗体の固定部
４ シンク
５ 試料液添加部
６ 基板
７ 標識付き抗体を保持するシート状多孔質担体
８ 検体を固定化する抗体を固定したシート状多孔質担
体
９ シンクを有するシート状多孔質担体1 Sheet-like porous carrier 2 Retaining part for labeled antibody 3 Immobilizing antibody immobilization part 4 Sink 5 Sample solution adding part 6 Substrate 7 Sheet-like porous carrier holding labeled antibody 8 Immobilizing antibody for immobilizing sample Sheet-like porous carrier 9 Sheet-like porous carrier having a sink
───────────────────────────────────────────────────── フロントページの続き (56)参考文献 特開 平１−227964（ＪＰ，Ａ) 特開 平８−15261（ＪＰ，Ａ) 特開 平５−10952（ＪＰ，Ａ) 特開 平６−341990（ＪＰ，Ａ) (58)調査した分野(Int.Cl.7，ＤＢ名) G01N 33/543 ─────────────────────────────────────────────────── ─── Continuation of front page (56) Reference JP-A-1-227964 (JP, A) JP-A-8-15261 (JP, A) JP-A-5-10952 (JP, A) JP-A-6- 341990 (JP, A) (58) Fields investigated (Int. Cl. 7 , DB name) G01N 33/543
Claims (2)
液を滴下または転写した後、風乾または凍結乾燥させる
ことのみからなるタンパク質の固定化方法であって、前
記シート状多孔質担体が、ガラス繊維、綿およびパルプ
からなる群より選ばれた少なくとも１つであることを特
徴とするタンパク質の固定化方法。1. A protein solution is dropped or transferred onto a sheet-shaped porous carrier and then air-dried or freeze-dried.
A method for immobilizing a protein consisting of the following, wherein the sheet-like porous carrier is at least one selected from the group consisting of glass fiber, cotton and pulp.
らなる群より選ばれた少なくとも１つである請求項１記
載のタンパク質の固定化方法。 2. The method for immobilizing a protein according to claim 1, wherein the protein is at least one selected from the group consisting of an enzyme, an antigen and an antibody .
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP10206497A JP3411470B2 (en) | 1997-04-18 | 1997-04-18 | Protein immobilization method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP10206497A JP3411470B2 (en) | 1997-04-18 | 1997-04-18 | Protein immobilization method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH10293127A JPH10293127A (en) | 1998-11-04 |
| JP3411470B2 true JP3411470B2 (en) | 2003-06-03 |
Family
ID=14317344
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP10206497A Expired - Fee Related JP3411470B2 (en) | 1997-04-18 | 1997-04-18 | Protein immobilization method |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP3411470B2 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5756611B2 (en) | 2010-10-07 | 2015-07-29 | 株式会社 資生堂 | Analysis method |
| CN105043056B (en) * | 2012-07-12 | 2017-10-31 | 艾博生物医药(杭州)有限公司 | A kind of method for automating flash baking sample reception pad |
-
1997
- 1997-04-18 JP JP10206497A patent/JP3411470B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JPH10293127A (en) | 1998-11-04 |
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