JPS589377B2 - electrophoresis analyzer - Google Patents

Description

[Detailed description of the invention] The present invention relates to an electrophoretic analysis device.

More specifically, the present invention relates to an electrophoretic analyzer equipped with a terminal electrolyte tank, a sample injection port, a migration tube with a detector interposed therein, a leading electrolyte tank, etc. The present invention relates to an electrophoresis analyzer in which at least one gel layer is provided in the leading electrolyte of an electrophoresis tube.

In the capillary type isotachophoresis analysis method, two types of electrolytes, a terminal electrolyte and a leading electrolyte, are filled inside the electrophoresis capillary, and substances that become charged at the interface between the two electrolytes, such as amino acids, A sample of peptides, biological substances, etc. is injected and subjected to constant current electrophoresis, and the target substance is separated into a single zone based on the difference in mobility, and qualitatively and/or quantitatively determined using a detector as appropriate. It is indispensable for trace analysis of samples as mentioned above.

However, recently, the targets of samples have become more diverse, and analysis of dilute samples or trace amounts of samples has come to be required, and the development of electrophoretic analyzers with even higher analytical accuracy has been anticipated.

The present invention was made in view of these circumstances, and one of its main structural features is that one end of the electrophoresis tube between the sample injection port and the detector of the electrophoresis analyzer is Alternatively, a gel layer for separation due to the difference in molecular weight is provided at both locations.

Due to this feature, the present invention can perform separation based on the difference in molecular weight in the gel layer (sieving effect) in addition to separation based on the difference in mobility, and can significantly improve the separation ability.

In other words, isokinetic electrophoresis analysis and zone electrophoresis analysis can be performed together with one device.

Furthermore, in electrophoresis using a gel layer, if a gradient layer with an appropriate pH is formed in the direction of electrophoresis, analysis by isoelectric focusing becomes possible, and the separation ability is further improved.

Preferred examples of the gel that can be used in the present invention to form the separation gel layer based on the difference in molecular weight include starch and agar.

Such a gel layer is formed at either end of the migration tube between the sample injection port and the detector, or at both locations.

Of course, a leading electrolyte layer is formed in a region other than the gel layer in the migration tube between the sample injection port and the detector.

As the electrophoresis tube in the present invention, in addition to those with an inner diameter of about 1 to 5 mmψ, capillary tubes (tubules) with an inner diameter of about 1 to 0.5 mmψ can be used, and in these cases, the provision of the gel layer part is For example, this can be done as follows.

In a thick tube (inner diameter 3 to 5 mmψ), the gelled material may be directly filled, but a predetermined amount of gel solution is placed in a predetermined part of the tube and photopolymerized using a daylight fluorescent lamp of about 10 W to form a gel. That's fine.

The present invention will be explained below based on the implementation sequence shown in the figures.

Note that the present invention is not limited to this.

In the figure, an isotachophoresis analyzer 1 includes a terminal electrolyte tank 2, a capillary tube 5 in which a sample injection port 3 and a potential gradient detector 4 are interposed, a leading electrolyte tank 6, and a sample injection port. A leading electrolyte layer section 7 and a gel layer section 8 are formed in the capillary tube 5 between the capillary tube 3 and the potential gradient detector 4.

Next, isotachophoresis analyzer 1 having the above configuration
Explain the operation.

First, an electrolytic solution containing anions with higher mobility than the sample (leading electrolyte) and an electrolytic solution containing anions with lower mobility (leading electrolyte) are placed at the position of the sample injection port 3 of the capillary tube 5.
A sample is injected into the interface, and electrophoresis is performed by supplying a constant current with a constant current high voltage power supply (not shown).

In this way, sample ions (anions, e.g. protein ions) are divided into zones (bands) of single component ions inside the capillary tube in the order of their mobility in the electrolyte tank 7.
The zone is separated into two zones, and each zone begins to move backward at a uniform speed with a constant width determined by the amount of ions while maintaining a clear boundary surface with each other.

These separation zones then enter the gel layer part 8, and are separated by the so-called "sieving effect," which is different from the separation due to differences in mobility in electrophoresis, and the separation zones described above are separated even more finely and clearly Become a thing.

For example, Figure 2 shows the differential value of the potential gradient in the electrolyte layer 7, but by passing through the gel layer 8, the fraction of P1 in Figure 2 is divided into two halves of Q1 and Q2 in Figure 3. It is expected that the separation accuracy will improve as the images are separated.

[Brief explanation of the drawing]

FIG. 1 is a functional explanatory diagram showing one row of the electrophoretic analyzer according to the present invention, FIG. 2 is a graph of the differential value of the potential gradient showing the state of separation of the target substance in the electrolyte tank, and FIG. The figure is a graph corresponding to Figure 2 showing the state of separation of the target substance in the gel layer. 1... Electrophoresis analyzer, 3... Sample injection port, 4...
...Detector, 5...Capillary tube, 8...Gel layer part.

Claims (1)

[Scope of Claims] 1. In an electrophoresis analyzer equipped with a terminal electrolyte tank, a migration tube with a sample injection port and a detector interposed therein, a leading electrolyte tank, etc. 1. An electrophoresis analyzer characterized in that a gel layer for separation based on a difference in molecular weight is provided at one or both ends of an electrophoresis tube. 2. The electrophoresis analyzer according to claim 1, wherein the electrophoresis tube is a thin tube.