JPH03172756A - Capillary electrophoretic device - Google Patents
Capillary electrophoretic deviceInfo
- Publication number
- JPH03172756A JPH03172756A JP1312907A JP31290789A JPH03172756A JP H03172756 A JPH03172756 A JP H03172756A JP 1312907 A JP1312907 A JP 1312907A JP 31290789 A JP31290789 A JP 31290789A JP H03172756 A JPH03172756 A JP H03172756A
- Authority
- JP
- Japan
- Prior art keywords
- capillary
- film
- immersed
- sample
- coated
- 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.)
- Granted
Links
- 238000013508 migration Methods 0.000 claims abstract description 5
- 230000005012 migration Effects 0.000 claims abstract description 5
- 238000005251 capillar electrophoresis Methods 0.000 claims description 5
- 239000004020 conductor Substances 0.000 claims description 3
- 229910052782 aluminium Inorganic materials 0.000 abstract description 14
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 abstract description 14
- 230000000694 effects Effects 0.000 abstract description 6
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 abstract description 6
- 239000000872 buffer Substances 0.000 abstract description 5
- 239000000463 material Substances 0.000 abstract description 5
- 239000007788 liquid Substances 0.000 abstract description 3
- 229910052697 platinum Inorganic materials 0.000 abstract description 3
- 230000005611 electricity Effects 0.000 abstract 1
- 230000008030 elimination Effects 0.000 abstract 1
- 238000003379 elimination reaction Methods 0.000 abstract 1
- 230000003068 static effect Effects 0.000 abstract 1
- 238000000576 coating method Methods 0.000 description 17
- 239000011248 coating agent Substances 0.000 description 15
- 238000001962 electrophoresis Methods 0.000 description 10
- 238000000034 method Methods 0.000 description 6
- 229910052751 metal Inorganic materials 0.000 description 5
- 239000002184 metal Substances 0.000 description 5
- 229920001721 polyimide Polymers 0.000 description 4
- 239000004642 Polyimide Substances 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 239000005350 fused silica glass Substances 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 238000005452 bending Methods 0.000 description 2
- 239000000428 dust Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000010306 acid treatment Methods 0.000 description 1
- 229920001222 biopolymer Polymers 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 238000005372 isotope separation Methods 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- -1 silver can be used Chemical compound 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
Landscapes
- Sampling And Sample Adjustment (AREA)
- Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)
Abstract
Description
【発明の詳細な説明】
（イ）産業上の利用分野
本発明は、タンパク質、核酸などの生体高分子の高分離
分析および精製法に係わり、特に構造や性質が接近した
成分の分離に好適なキャピラリ電気泳動装置に関する。Detailed Description of the Invention (a) Industrial Application Field The present invention relates to high-level separation analysis and purification methods for biopolymers such as proteins and nucleic acids, and is particularly suitable for separating components with similar structures and properties. This invention relates to a capillary electrophoresis device.
（ロ）従来技術
電気泳動の窩性能装置化技術として、近年急速に研究が
なされているキャピラリ電気泳動法は、Ｊ、 Ｌ Ｊｏ
ｒｇｅｎｓｏｎやＢ、 Ｌ、 Ｋａｒｇｅｒらの総説（
各々、５ｃｉｅｎｃｅ、２２２２６６（１９８３） Ｊ
ｏｕｒｎａｌ ｏｆ Ｃｈｒｏｍａｔｏｇｒａ−ｐｈｙ
、虱５８５（１９８９）、 ）において示されているよ
うに、１）キャピラリから効果的にジュール熱を除去す
ることによって高圧で泳動ができるため、原理的に高分
離能、迅速分析が可能、２）オンカラム検出器によって
装置の自動化が可能等の利点を有しペプチド、タンパク
質、核酸の分前分析及び精製をはじめとして、光学分割
、同位体の分離その他極めて酷似した成分間の分離に適
した方法である。(b) Conventional technology The capillary electrophoresis method, which has been rapidly researched in recent years as a device-based technology for electrophoresis, is the method described by J. L. Jo.
Reviews by Rgenson, B. L., Karger et al.
5science, 222266 (1983) J
internal of chromatography
, 585 (1989), ), 1) Since Joule heat is effectively removed from the capillary, electrophoresis can be performed at high pressure, which in principle enables high resolution and rapid analysis; 2) ) A method suitable for preparative analysis and purification of peptides, proteins, and nucleic acids, as well as optical resolution, isotope separation, and other separations between extremely similar components, with advantages such as the ability to automate the device using an on-column detector. It is.
なお、キャピラリには通常、フユーズドシリカが使用さ
れ、その曲げ強度を増大するために、キャピラリ外面に
ポリイミド被膜をコーティングしている。Note that fused silica is usually used for the capillary, and the outer surface of the capillary is coated with a polyimide film to increase its bending strength.
（ハ）発明が解決しようとする課題
しかしながら、ポリイミド被膜は非常に熱伝以下 余白
導が悪い究めキャピラリの外側から温調を行う場合に効
果的な温調を行えなかった。(c) Problems to be Solved by the Invention However, the polyimide coating has extremely low heat conductivity and poor margin conductivity, making it impossible to effectively control the temperature from the outside of the capillary.
また、キャピラリ両端ＫＩＯ〜５０ＫＶを印加して電気
泳動を行うが、ポリイミド被膜は非導電性のためキャピ
ラリ内部と周辺との電位差による静電的影響を生じてい
た。これは、具体的には光学検出器の検出部（フォトセ
ル）、検出回路に対する静電誘導、ホコリ等をひきつけ
ることにより光路全妨げるために生じるノイズ等である
。Furthermore, electrophoresis is performed by applying KIO to 50 KV to both ends of the capillary, but since the polyimide coating is non-conductive, electrostatic effects occur due to the potential difference between the inside and the surroundings of the capillary. Specifically, this includes electrostatic induction to the detection section (photocell) of the optical detector and the detection circuit, and noise that occurs because it attracts dust and the like and obstructs the entire optical path.
そこで１本発明は、キャピラリの温調を効果的に行え、
かつ、静電的影響を除去した装置を提供することケ目的
とする。Therefore, one aspect of the present invention is to effectively control the temperature of the capillary.
In addition, it is an object of the present invention to provide a device that eliminates electrostatic influences.
に）課題を解決する友めの手段
本発明は、上記課題を解決するｔめ１曲げ強度を増大す
るためのコーティングに熱伝導の良い材質を用いること
を特徴とする〇
ここで、熱伝導の良い材質とは１例えば、アルミ、金、
銀などの金属を挙げることができ、特にアルミが好まし
い。B) A companion means for solving the problems The present invention is characterized in that a material with good thermal conductivity is used for the coating for increasing the bending strength. Good materials are 1.For example, aluminum, gold,
Metals such as silver can be used, and aluminum is particularly preferred.
かかる材質ヲキャビラリにコーティングして被膜をｆ′
ｆ：ｆＭする方法としては９例えば、金属を溶融して、
へヶで塗る方法、スプレーで吹きつける方法、金属の溶
融液にキャピラリヲつける方法など何でも良い。Such a material is coated on the cavity to form a film f′.
f: As a method for fM, 9 For example, by melting metal,
Any method is suitable, such as applying it with a spatula, spraying it, or dipping a capillary into the molten metal.
なお、熱伝導性材質をコーティングしたキャピラリを泳
動管として用いる場合には、キャピラリ両端の泳動用緩
衝液と接する部分のコーティングをはぎとることが必要
である。Note that when a capillary coated with a thermally conductive material is used as an electrophoresis tube, it is necessary to strip off the coating on the portions of the capillary that are in contact with the electrophoresis buffer at both ends.
また１本発明では、キャピラリ内部と周辺との電位差に
よる静電的影響を除去するため、上記熱伝導材質でコー
ティングした泳動管の一部を接地することを特徴とする
。Further, the present invention is characterized in that a part of the migration tube coated with the heat conductive material is grounded in order to eliminate the electrostatic influence due to the potential difference between the inside of the capillary and its surroundings.
（ホ）作用
本発明では、熱伝導性良好な材質の被膜によりキャピラ
リのほぼ全体がすきまなく密着しているため、被膜の温
調によシ効果的な温調が実現される。(E) Function In the present invention, since almost the entire capillary is tightly adhered to the capillary without any gaps by the coating made of a material with good thermal conductivity, effective temperature control can be achieved by controlling the temperature of the coating.
また、接地することＫよシ金属被膜がシールド効果を持
ち静電的影響を除去できる。Furthermore, since it is grounded, the metal coating has a shielding effect and can eliminate electrostatic influences.
（へ）夾施例
本発明に部るキャピラリ電気泳動装置の概略図の一実施
例を第１図に示す。(F) Additional Examples An example of a schematic diagram of a capillary electrophoresis apparatus according to the present invention is shown in FIG.
図中１は、フユース゛ドシリカキャピラリ（内径１０〜
１００μｍ、長さ１０ｃＭ〜１ｍ）で、アルシミ被暎１
を施しており、その両端］Ａ、ｌＢは、酸処理により部
分的に被膜がはぎとっである。なお、かかるＩＡ 、
ＩＢの拡大図を第２図に示しである。゛被膜１′をはぎ
とったＩＡ 、 ＩＢは泳動用緩衝液槽７゜８につけら
れる。1 in the figure is a fused silica capillary (inner diameter 10~
100μm, length 10cM~1m), aluminum resistant 1
The coating at both ends A and IB was partially peeled off due to acid treatment. In addition, such IA,
An enlarged view of IB is shown in FIG. IA and IB from which the coating 1' has been removed are placed in an electrophoresis buffer tank 7.8.
アルミ被膜１′ヲ施しであるキャピラリは、アルミブロ
ック２にはさみこまれ、このブロック２はベルチェ素子
３により温調される。A capillary coated with an aluminum coating 1' is inserted into an aluminum block 2, and the temperature of this block 2 is controlled by a Bertier element 3.
４は、アルミブロック２の温度を検出するための温度セ
ンサ（例えば、抵抗・ダイオード等）５は慇度センサ４
の検出回路及びベルチェ素子の温調用回路である。4 is a temperature sensor (for example, a resistor, a diode, etc.) for detecting the temperature of the aluminum block 2; 5 is a temperature sensor 4;
This is a detection circuit and a temperature control circuit for the Beltier element.
また、６は電気泳動を行うための高圧電源で最大出力電
圧３０ ＫＶ程度のものが好ましり、電源６には、白金
型（至）１０が接続される。Further, 6 is a high-voltage power source for performing electrophoresis, preferably having a maximum output voltage of about 30 KV, and a platinum mold 10 is connected to the power source 6.
９は、検出器で例えばＵＶ検出器あるいは螢光検出器が
用いられる。Reference numeral 9 denotes a detector, such as a UV detector or a fluorescence detector.
以上の構成において動作は次の様に行う。In the above configuration, the operation is performed as follows.
まず、キャピラリ１の−＠を、試料容器（図示せず）に
浸しキャピラリ１内に試料全導入する。First, -@ of the capillary 1 is immersed in a sample container (not shown), and the entire sample is introduced into the capillary 1.
試料導入後、キャピラリ１の両端全泳動用緩衝液を入れ
た緩衝液槽７．８に浸す。この二つの緩僅工液槽に各々
白金電嘩ｌＯを浸し両甑に電圧を印加する。このとき、
キャピラリにジュー／Ｌ／Ｐ：が発生するため、それを
抑えるためアルミブロック２により冷却する。After introducing the sample, both ends of the capillary 1 are immersed in a buffer tank 7.8 containing a buffer solution for electrophoresis. A platinum electric wire 1O is immersed in each of these two slow-working liquid tanks, and a voltage is applied to both pots. At this time,
Since juice/L/P: is generated in the capillary, it is cooled by an aluminum block 2 to suppress it.
キャピラリ両端に電Ｅｅ印加することによって。By applying an electric current Ee to both ends of the capillary.
緩＃液に流れが生じ、試料が分離され１分港された試料
成分は検出器９で検出される。A flow is generated in the slow liquid, the sample is separated, and the sample components that have been left for 1 minute are detected by the detector 9.
以上の動作において２本発明ではキャピラリの被膜ニア
ルミを用いているため、キャビフリ表面が効果的にかつ
均一に温調される。In the above-mentioned operation, the present invention uses a capillary coating made of aluminum, so the temperature of the surface of the capillary can be effectively and uniformly controlled.
なお、アルミ被膜ヲ施したキャピラリの強度は両者の熱
膨張率が異なるため、低温あるいは高温での使用には問
題があるが、電気泳動は０通常室温付近で行うため問題
は生じない。It should be noted that the strength of the capillary coated with aluminum is problematic when used at low or high temperatures because the coefficients of thermal expansion are different, but this problem does not occur because electrophoresis is usually performed at around room temperature.
また１本発明の他の実施例を第３図に示す。第１図と同
じものには同じ番号が付してあり、この実施例では、ア
ルミ被膜１′の一部が接地されている（図中１１）。か
かる接地により、高電圧印加部のは＃了全域で被膜によ
る静電シールドをツ現できる。Another embodiment of the present invention is shown in FIG. Components that are the same as those in FIG. 1 are given the same numbers, and in this embodiment, a portion of the aluminum coating 1' is grounded (11 in the figure). This grounding allows electrostatic shielding by the film to be achieved over the entire area of the high voltage application section.
（ト）効果
本発明によれば、キャピラリ表面が効果的にかつ均一に
温調され高分解能の分離を短時間で実現すると共に再現
性が向上する。(g) Effects According to the present invention, the temperature of the capillary surface is effectively and uniformly controlled, high-resolution separation is realized in a short time, and reproducibility is improved.
また、被膜外部との接触が不完全（一部接触していない
部分がある時）な場合、従来のポリイミド被膜では局部
的に温調されないが１例えばアルミ被膜では熱伝導率が
上記被膜の５００〜１０００倍であるので、かかる事態
が大幅に緩和される。In addition, when the contact with the outside of the coating is incomplete (when there are some parts that are not in contact), the temperature cannot be controlled locally with conventional polyimide coatings.1 For example, with aluminum coatings, the thermal conductivity is 500 Since it is 1000 times larger, such a situation can be greatly alleviated.
更に、金属被膜のシールド効果により、静電誘導、ホコ
リのすい込みが防止される。Furthermore, the shielding effect of the metal coating prevents electrostatic induction and dust from entering.
第１図、第３（２）は１本発明に係る電気泳動装置の一
夾施例図、第２図は、キャピラリの両端の状態を示す図
である。
ｌ・・・フユーズドシリカキャヒ′ラリ】・・・アルミ
被膜FIGS. 1 and 3(2) are views of one embodiment of an electrophoresis device according to the present invention, and FIG. 2 is a view showing the states of both ends of a capillary. l...Fused silica capillary]...Aluminum coating
Claims (1)
を泳動管として用いてなるキャピラリ電気泳動装置。 ２、請求項第１項の泳動管の一部を接地してなるキャピ
ラリ電気泳動装置。[Claims] 1. A capillary electrophoresis device using a capillary whose outer surface is coated with a thermally conductive material as a migration tube. 2. A capillary electrophoresis device in which a part of the migration tube according to claim 1 is grounded.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1312907A JPH0737962B2 (en) | 1989-11-30 | 1989-11-30 | Capillary electrophoresis device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1312907A JPH0737962B2 (en) | 1989-11-30 | 1989-11-30 | Capillary electrophoresis device |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH03172756A true JPH03172756A (en) | 1991-07-26 |
| JPH0737962B2 JPH0737962B2 (en) | 1995-04-26 |
Family
ID=18034893
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1312907A Expired - Fee Related JPH0737962B2 (en) | 1989-11-30 | 1989-11-30 | Capillary electrophoresis device |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0737962B2 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5180475A (en) * | 1991-09-04 | 1993-01-19 | Hewlett-Packard Company | System and method for controlling electroosmotic flow |
| US5322607A (en) * | 1992-07-14 | 1994-06-21 | Hewlett-Packard Company | Electrical potential configuration for an electrophoresis system |
| US6103081A (en) * | 1996-12-11 | 2000-08-15 | The Regents Of The University Of Michigan | Heat sink for capillary electrophoresis |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6168561A (en) * | 1984-09-12 | 1986-04-08 | Fujikura Ltd | Capillary column for chromatograph |
-
1989
- 1989-11-30 JP JP1312907A patent/JPH0737962B2/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6168561A (en) * | 1984-09-12 | 1986-04-08 | Fujikura Ltd | Capillary column for chromatograph |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5180475A (en) * | 1991-09-04 | 1993-01-19 | Hewlett-Packard Company | System and method for controlling electroosmotic flow |
| US5322607A (en) * | 1992-07-14 | 1994-06-21 | Hewlett-Packard Company | Electrical potential configuration for an electrophoresis system |
| US6103081A (en) * | 1996-12-11 | 2000-08-15 | The Regents Of The University Of Michigan | Heat sink for capillary electrophoresis |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0737962B2 (en) | 1995-04-26 |
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| Date | Code | Title | Description |
|---|---|---|---|
| LAPS | Cancellation because of no payment of annual fees |