JPH06226054A - Electrophoresis device for continuous treatment - Google Patents

Abstract

PURPOSE:To realize the device which can be continuously operated and prevents the inclusion of air bubbles into a migration cell, etc., by decreasing the amt. of the electrolyte soln. to be used and eliminating the need for an electrolyte soln. tank of a large capacity. CONSTITUTION:This electrophoresis device has an electrolyte soln. regeneration treating device 6 which is provided between the migration cell 3 to be injected with a sample from a sample injector 2 and to be supplied with the electrolyte soln. from the electrolyte soln. tank 1 and the electrolyte soln. tank 1 via an electrolyte soln. feed pump 4 and has a separating membrane 7. As a result, the separating membrane 7 removes a slight amt. of the sample and, therefore, the reutilization of the electrolyte soln. after the use is possible and the need for prepn. of a large amt. of the electrolyte soln. and the electrolyte soln. tank 1 of the large capacity is eliminated. Since an electrolyte soln. piping is formed as a closed loop, a continuous automatic operation is possible. The possibility of the intrusion of the air bubbles is eliminated and, therefore, the electrophoresis maintaining sepn. accuracy is possible.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an electrophoretic device for continuous processing for continuously processing a sample.

[0002]

2. Description of the Related Art When an electric field is applied to a sample having a charge in an electrolyte solution such as protein or cell, these samples perform electrophoresis. Since the speed of electrophoresis in a solution varies depending on the amount of electric charge of each sample, electrophoresis is used for separation, purification, identification, etc. of these samples.

In the conventional electrophoresis apparatus, the electrolyte solution used is stored in the electrolyte solution tank 1 as shown in FIG. 3, and after being subjected to the electrophoresis, a small amount of the solution containing a large amount of the sample is stored. Although stored in the sorting container 5, most of the electrolyte solution containing almost no sample is directly collected in the waste liquid container 8 and discarded.

Therefore, if all the electrolyte solution in the electrolyte solution tank 1 is used up, it is necessary to replace the electrolyte solution tank 1, replenish the electrolyte solution, or change the connection of the electrolyte solution pipes. I have to stop.

[0005]

In the conventional device,
The following problems were encountered when trying to carry out electrophoresis continuously. (1) A large amount of electrolyte solution must be prepared, a large electrolyte solution tank is required, and the device becomes large. (2) When the remaining amount of the electrolyte solution in the electrolyte solution tank becomes very small, it is necessary to replace the electrolyte solution tank, replenish the electrolyte solution, or reconnect the electrolyte solution pipes, and stop the electrophoresis. There must be. (3) At the time of work such as reconnection of the above-mentioned pipes, bubbles may enter the pipes or the migration tank, which may lead to a decrease in separation accuracy. The present invention is intended to solve the above problems.

[0006]

An electrophoretic device for continuous processing according to the present invention is provided with an electrophoretic tank having an anode electrode and a cathode electrode therein, an electrolyte solution tank for supplying an electrolytic solution to the electrophoretic tank, and a sample. In an electrophoretic device equipped with a sample injector and a preparative container from which the preparative liquid is discharged from the electrophoretic tank, the electrophoretic solution feed pump is connected between the electrophoretic tank and the electrolytic solution to separate the electrophoretic solution into the inside. It is characterized in that it is provided with an electrolyte solution regeneration treatment device provided with a membrane.

[0007]

In the above, the sample injected from the sample injector into the electrophoretic solution tank supplied with the electrolyte solution is subjected to electrophoresis, separation, purification, identification, etc., and then many samples are added. The contained electrolyte solution is dispensed in a dispensing container.

The uncollected electrolyte solution is sent to the electrolyte solution regeneration treatment device by the electrolyte solution sending pump,
Since the separation membrane provided in the device separates most of the sample and allows small electrolyte solution ions to pass through, the above regeneration treatment device has basic properties that greatly affect the electrophoresis such as the pH and conductivity of the electrolyte solution. Collect the unchanged electrolyte solution and supply it to the electrolyte solution tank.

As described above, after the electrolyte solution regeneration treatment device removes a small amount of sample contained in the electrolyte solution after electrophoresis, it is returned to the electrolyte solution tank and can be reused for electrophoresis. There is no need to prepare the above electrolyte solution, and a large electrolyte solution tank is not required. Also, because the electrolyte solution piping is a closed loop,
Samples can be processed continuously by automatic operation, and
Since there is no risk of air bubbles entering the piping or the migration tank,
Continuous electrophoresis is possible while maintaining the separation accuracy.

[0010]

FIG. 1 shows an embodiment of the present invention. In this embodiment shown in FIG. 1, an electrophoretic bath 3 having an anode electrode 3a and a cathode electrode 3b therein, a sample injector 2 for injecting a sample into the electrophoretic bath 3, and an electrolyte solution are supplied to the electrophoretic bath 3. An electrophoretic device comprising an electrolyte solution tank 1, an electrolyte solution feed pump 4 connected to the tank 1, and a sorting container 5 to which a fractionation liquid 5a is fed from the migration tank 3, wherein: An electrolyte solution regeneration treatment device 6 that is connected between the electrolyte solution feed pumps 4 and has a separation membrane 7 inside is provided.

In the above, in the migration tank 3 in which the electrolyte solution is supplied from the electrolyte solution tank 1 and the sample is injected by the sample injector 2, the sample is electrophoresed between the electrodes 3a and 3b to separate the sample. After purification, identification, etc.,
The electrolyte solution containing a large amount of the sample is collected in the collecting container 5.

The uncollected electrolyte solution is sent to the electrolyte solution regeneration treatment device 6 by the electrolyte solution feed pump 4. There is a separation membrane 7 in the electrolytic solution regeneration processing device 6, in which the sample component and the electrolytic solution are separated, and the electrolytic solution is regenerated. The regenerated electrolyte solution is returned to the electrolyte solution tank 1 and again guided to the migration tank 3 for reuse.

Regeneration of the electrolyte solution in the electrolyte solution regeneration treatment device 6 is carried out by separating the sample with the separation membrane 7. However, the size of the protein, which is a typical sample for electrophoresis, is several tens of angstroms. , Cells have tens of thousands of angstroms, whereas the ions that make up the electrolyte solution are as small as a few angstroms, the separation membrane 7 can separate most of the sample from the electrolyte solution, and small electrolyte solution ions can pass through. Therefore, the basic properties such as pH and conductivity of the electrolyte solution, which greatly affect electrophoresis, do not change.

In order to confirm changes in the basic properties of the electrolyte solution, human serum albumin was used as a sample, and 7 mM triethanolamine / acetic acid (pH 7.5 at 2) was used as the electrolyte solution.
0 ° C), UF membrane as a separation membrane (molecular weight cut-off: 10,000)
0) was used to perform the experiment.

From the results of this experiment, it can be seen that, as shown in FIG. 2, even if the electrophoresis and the regeneration treatment are repeated at least four times or more, the basic properties important for electrophoresis, pH and conductivity, do not change. confirmed.

From the above, it has become possible to continuously perform electrophoresis for four times or more without increasing the size of the electrolyte solution tank. In addition, if the same amount of sample is processed, the electrolyte solution can be processed with a quarter or less of the required amount, so that the capacity of the electrolyte solution tank can be reduced to a quarter.
The following can be achieved, and the device can be downsized.

On the other hand, since the electrolyte solution pipe can be closed loop and the sample can be continuously processed by almost automatic operation, there is no possibility that bubbles enter the pipe or the migration tank, and the separation accuracy is maintained. It became possible to process the sample continuously in the state.

[0018]

The electrophoretic device for continuous processing of the present invention includes an electrolyte solution feed pump between the electrophoresis tank in which the sample is injected from the sample injector and the electrolyte solution is supplied from the electrolyte solution tank, and the electrolyte solution tank. Since the separation membrane removes a trace amount of the sample, it is possible to reuse the electrolyte solution after use by preparing an electrolyte solution regeneration treatment device having a separation membrane provided through A large-capacity electrolyte solution tank is no longer required, and the electrolyte solution piping forms a closed loop, which allows continuous automatic operation and eliminates the possibility of air bubbles being mixed in, enabling electrophoresis that maintains separation accuracy.

[Brief description of drawings]

FIG. 1 is an explanatory diagram of a continuous electrophoresis device according to an embodiment of the present invention.

FIG. 2 is an explanatory diagram of a change in basic properties of the electrolyte solution according to the above embodiment after the regeneration treatment, in which (a) is a change in pH and (b) is an explanatory view of a change in conductivity.

FIG. 3 is an explanatory diagram of a conventional electrophoresis device.

[Explanation of symbols]

 1 Electrolyte Solution Tank 2 Sample Injector 3 Migration Tank 3a, 3b Electrode 4 Electrolyte Liquid Delivery Pump 5 Preparative Container 5a Preparative Liquid 6 Electrolyte Solution Regeneration Device 7 Separation Membrane

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Internal reference number FI technical display location G01N 27/447

Claims (1)

[Claims] 1. A migration tank in which an anode electrode and a cathode electrode are provided, an electrolyte solution tank for supplying an electrolyte solution to the migration tank, a sample injector for injecting a sample, and a preparative liquid discharged from the migration tank. In an electrophoretic device having a preparative separation container, an electrolytic solution regeneration treatment device is provided which is connected between the electrophoretic bath and the electrolytic solution via an electrolytic solution feed pump and has a separation membrane provided therein. Characteristic electrophoretic device for continuous processing.