JPS60138447A - Electrophoresis apparatus - Google Patents

Abstract

PURPOSE:To prevent an electroosmosis flow and an interference between an electroosmotic agent and a sample by introducing an electrolyte to which the electroosmotic agent is added, into an electrophoresis tube, then introducing the electrolyte to which the electroosmotic agent is not added. CONSTITUTION:A tube 8 for supplying the electrolyte containing the electroosmosis preventing agent and a tube 9 for supplying the electrolyte containing no said agent are connected to one end of the electrophoresis tube 5 through a switching valve 10. Under the state of the valve 10 as shown in a figure, a stroke pump 15 is driven to wash the inside of the tube 5 with the electrolyte containing electroosmosis preventing agent. Next, the valve 10 is changed over, and the pump 15 is driven similarly to introduce a sample liquid (mixed liquid of the sample and the electrolyte containing no electroosmosis preventing agent) into the tube 5 from the tube 9 while pushing away the electrolyte containing said agent. Thereafter, operation of the pump 15 is stopped, and the electrophoresis is started. The electrophoresis velocity of the sample is detected by a scattered light detector 4.

Description

Detailed Description of the Invention (a) Industrial Application Field This invention relates to an electrophoresis device, and particularly an improvement to avoid interference between an inhibitor mixed into an electrolytic solution to prevent electroosmotic flow and a sample. Concerning configuration.

(b) Conventional technology Conventionally, electroosmotic flow (Electr.

To avoid the influence of osmotic flow, Trito/Hydroxypropyl methyl cellulose (rM
o), electroosmotic inhibitors such as polyvinyl alcohol (PV) are used mixed into the electrolyte. However, if these electroosmotic inhibitors are mixed in the sample, it is possible that they may cause some kind of interference with the sample.

In particular, Triton is a type of surfactant, and when the sample is a cell, it causes a change in the charge of the cell.

On the other hand, it has been proposed to coat the inner surface of the electrophoresis tube with an electroosmotic inhibitor, but this has problems with durability (see Japanese Patent Laid-Open No. 57-30940).

(C) Purpose This invention was made in view of these circumstances, and one of its main purposes is to provide an electrophoresis device that can prevent interference of an electroosmotic inhibitor with a sample.

(Two configurations) This invention has a pair of electrode vessels, and connects these electrode vessels.
An electrophoresis path equipped with a detector is provided, and an electrolytic solution supply path containing an electroosmosis inhibitor and an electrolytic solution supply path not containing an electroosmosis inhibitor are connected near one end of this migration path via a switching valve. This is an electrophoresis device in which an electrophoresis channel is connected to the electrophoresis channel, and an electrolyte drain channel is connected near the other end of the migration channel.

(e) Examples The present invention will be described in detail below based on examples shown in the drawings. Note that this invention is not limited to this.

In Fig. 1, a cell electrophoresis device (1) has a pair of electrode tanks (2) and (3), and a detector (4) that connects both electrode tanks.
), and furthermore, this wave tube is equipped with an electrophoresis tube (5) equipped with an electrophoresis tube (5) having an electrolytic solution containing an electrolytic osmosis inhibitor and a sample solution in which a sample is mixed with an electrolytic solution containing no electrolytic osmotic inhibitor on the inlet side. A switching supply means (6) t and a drainage means (7) are provided on the outlet side.

The switching supply means (6) connects an electrolyte supply pipe (8) containing an electroosmosis inhibitor and a sample solution supply pipe (9) containing an electrolyte containing a sample and an electroosmosis inhibitor. Switching valve α
It is connected to the switching supply pipe (b) via Q. Incidentally, ``O'' is a sample liquid container, and ``O'' is an electrolytic solution container that does not contain an electroosmotic inhibitor.

On the other hand, the liquid draining means (7) consists of a liquid draining pipe α and a draining pump α which also serves as a supply power source for the switching supply means (6). In addition, (g) is a drainage container, and qη (g) is a diaphragm.

Next, the operation of the cell electrical turbulence device (1) having the above configuration will be explained.

First, set the switching valve Q13 to the state shown in Figure 1, and then set the straining pump (
When the electrophoresis tube (5) is activated, an electrolytic solution containing an electroosmotic inhibitor (to be added) flows through the turbulent tube (5) and cleans it.

Next, when switching the switching valve head, the same squeezing pump Q0
As a result of the operation, a mixed solution of the sample solution, a pickled sample, and an electrolytic solution (wave buffer) that does not contain an electroosmotic inhibitor (not added) flows into the turbulent tube (5), and an electrolytic solution containing an electroosmotic inhibitor is mixed with the electrolytic solution that contains an electroosmotic inhibitor. Note that compartment air may be introduced before introducing the sample liquid into the turbulence tube.

In this way, the straining pump α time is stopped and electrophoresis is started. The electrophoresis speed of the sample is measured by irradiating the sample during electrophoresis with light and detecting the scattered light with the detector (4).A zero measurement result indicates that the electrolytic solution used substantially contains an electroosmotic inhibitor. The effects of electroosmosis are removed, even though they are not present.

This is because before the sample solution is supplied into the electrophoresis tube (5), an electrolytic solution containing an electroosmotic inhibitor is supplied, and this electrolytic solution remains in a coating state on the inner surface of the electrophoresis tube (5). It is thought that this is because of this. Of course, measurement can be performed without changing the amount of charge on the sample.

After electrophoresis, switch the switching valve (10) again and press the
Activate f9 to wash the inside of the electrophoresis tube (5) with an electrolytic solution containing an electroosmotic inhibitor. Note that this washing requires 50% of the sample solution.
It is preferable to use a batch amount of electrolyte.

Further, instead of washing each time the electrophoresis measurement is performed as described above, the washing may be performed after repeating the electrophoresis measurement multiple times, for example, up to about 10 times.

Here, for reference, we will show an example of an experiment conducted using a capillary type constant-velocity electric wave device.

Leading electrolyte e: Morpholinoethanesulfonic acid [5 mmol] 2-amino-2-methyl-1,3-propanediol (amediol) (10 mmol) Terminal electrolyte: Ibsilone-aminocaproic acid [5 mmol] 2-amino 2-Methyl-1,3-propanediol (Amediol) and Ba (OR) g [: 10 mmol] Electrophoresis current: 150 μm (9 minutes no (change 2 → 75 μm permeation tube: 11/X4σ, detector: Potential gradient detector sample:
Organic acids (A, B, O, D) Electrolyte used: ■No electroosmotic agent added, ■Electroosmotic agent added, Hydroxypropylmethyl (EFMO) (0.1% by weight) Electrolytic solution with electroosmotic agent added After introducing the electrolyte, the same additive-free electrolyte as in ■ was introduced without displacing the electrolyte. The electropherograms of experimental results ■, ■, and ■ are shown in Figures 2, 3, and 4, respectively. Figure 4 shows that if an electrolytic solution containing an electroosmotic agent is introduced in advance, the effect of electroosmosis can be prevented even if the added electrolytic solution is subsequently pushed away and an electrolytic solution without an electroosmotic agent is introduced for electrophoresis. I understand.

(Hen Effect) This invention prevents electroosmotic flow and prevents electroosmotic flow by configuring the electrophoresis tube so that an electrolytic solution containing no electroosmotic agent can be introduced after introducing an electrolytic solution containing an electroosmotic agent. It is also possible to prevent sample interference.

[Brief explanation of drawings]

Fig. 1 is a functional explanatory diagram showing an embodiment of the electric wave device according to the present invention, Fig. 2 is an electrophoresis diagram when the electrolyte is not added with an electroosmotic agent, and Fig. 3 is an electrophoresis diagram when the electrolytic solution is added with an electroosmotic agent. Figure 4 shows the electropherogram of the case after using electrolyte with electroosmotic agent added.
FIG. 2 is an electropherogram when an electrolytic solution containing no electroosmotic agent is used. (1)・・・・・・Cell electrophoresis device, (2)(3)・
... Electrode tank, (4) ... Detector, (5
)...Transfer tube, (7)... Drainage means, (8)... Electrolyte supply pipe containing an electroosmotic agent,
(9)... Sample liquid supply tube, <IQ...
・Switching valve.

Claims (1)

[Claims] 1. Equipped with a pair of electrode tanks and an electrophoresis path that connects these electrode tanks and is equipped with a detector, and further includes an electrolytic solution supply path containing an electroosmosis preventive agent and an electroosmosis preventive agent near one end of this migration path. Connect the electrolyte supply path that does not contain the agent through the switching valve,
An electrophoresis device comprising an electrophoresis channel connected to the other end of the electrophoresis channel.