JPS60250243A - Uniform velocity electrophoretic apparatus - Google Patents

Abstract

PURPOSE:To enable the lowering of an electrophoretic voltage by providing an electrode moving means (or an electrophoretic tube moving means) for bringing the electrode in a terminal electrolytic liquid tank close to a potential gradient detector and a voltage detector for monitoring an electrophoretic voltage. CONSTITUTION:An electrode 21 in a terminal electrolytic liquid tank 2 is moved horizontally in the axial direction of an electrophoretic tube 4 through a liquidtight packing 23 while an electrode moving section 22 is provided having a rack pinion mechanism and a stepping motor. Also a control section 51 is provided which has a voltage detector for monitoring a voltage between poles of electrodes 21 and 31. Then, when the detection voltage exceeds a specified value, a signal is sent to the electrode moving section 22 to move the electrode 21 gradually to a potential gradient detector 42 whereby it is controlled to keep the detection voltage from exceeding the value. Thus, the capacity of an electrophoretic power source, especially the maximum voltage can be lowered by holding down the electrophoretic voltage requiring a high voltage.

Description

DETAILED DESCRIPTION OF THE INVENTION (a) Field of Industrial Application This invention relates to an isokinetic electrophoresis device. More specifically, the present invention relates to an isotachophoresis device that improves the electrophoresis voltage between the two electrodes during electrophoresis.

(2)) Conventional isotachophoresis analysis method involves filling a migration tube with a terminal electrolyte and a leading electrolyte, and placing a substance that can be charged at the interface between the two electrolytes, such as amino acids, peptides, etc.
This is a method in which a sample such as a biological substance 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 qualitative and quantitative analysis is performed using a detector as appropriate. This method is suitable for microanalysis of large samples.

However, when electrophoresis is performed with the intention of such analysis in a conventional isotachophoresis apparatus, the terminal electrolyte zone with low conductivity increases between the two electrodes as the electrophoresis progresses, so the voltage between the two electrodes often reaches a high value ( For example, 20KV)
shows. Therefore, the capacity of the power supply for electrophoresis must be increased, and the base voltage becomes high, making it difficult to separate and detect the target sample zone.In addition, there are many problems such as the need to improve the insulation of the entire system. There was a problem.

e) Purpose This invention was made in view of the above situation,
The present invention aims to provide an isokinetic electrophoresis device that can reduce the electrophoresis voltage.

B) Structure According to the present invention, there is provided an isokinetic electrophoresis device in which a terminal electrolyte tank and a leading electrolyte tank are connected by a migration tube, and the migration tube is provided with a sample injection port and a potential gradient detector. The terminal is equipped with electrode moving means or electrophoresis tube moving means for moving the electrode set in the terminal electrolyte tank close to the potential gradient detection type, and a voltage detector for monitoring the electrophoresis voltage during electrophoresis. There is provided a constant-velocity electrophoresis apparatus which is configured to operate the moving means to prevent or reduce the electrophoresis voltage when the voltage exceeds a predetermined value.

The predetermined value of the electrophoresis voltage and IJ mean a test pressure value that is lower than the electrophoresis voltage that allows electrophoresis analysis to be performed at least without operating the electrode or the electrophoresis tube moving means, and the performance of the potential gradient detector, the capacity of the power supply, etc. It is appropriate to set the value as low as possible within the allowable voltage range. Usually, by adjusting the moving distance of the electrodes and the migration tube, it is possible to set the voltage to a value that cuts 70 to 80% of the maximum electrode-to-electrode voltage required in conventional devices.

(e) Examples The isokinetic electrophoresis apparatus of the present invention will be described in detail below using examples.

(IA) shown in FIG. 1 is a configuration explanatory diagram showing an embodiment of the isotachophoresis apparatus of the present invention. In the figure, the electrophoresis apparatus (IA) consists of a terminal electrolyte tank (2) and a leading electrolyte tank (3) connected by a migration tube (4). Entrance (41
), but there is a potential gradient detector (4) on the leading electrolyte tank side.
2 is provided. Then, the terminal electrolyte tank (2
Electrodes e2]1.4'l11 are installed in the + and leading electrolyte tanks (3), respectively, and these are connected to a constant current DC power supply (5) and set so that an electrophoresis voltage can be applied. However, the terminal electrolyte tank (electrolyte PjA 1 in 21) can be moved horizontally along the axis of the migration tube (4) via a liquid-tight gasket (B), and can be mounted on the rack. It is equipped with a pinion mechanism and a stepping motor, and is supported in two or five pieces.

Further, (51) is a control unit equipped with a voltage detector that monitors the voltage between the two electrodes, and when this detected voltage exceeds a predetermined value (for example, 4 KV), it sends a signal to the electrode moving unit (22). ) is gradually moved in the direction of the potential gradient detector to perform feedback control so that the detected voltage does not exceed a predetermined value.

The operation of such an isotachophoresis apparatus will be explained below.

First, Figure 4 shows the change over time in the state of the zone inside the migration tube during isokinetic electrophoresis, where A is the start of electrophoresis, σ) is the terminal electrolyte, (S) is the injected sample, ( L) means leading electrolyte. As electrophoresis progresses, the zones become B and C! After passing through fM, the state becomes D, and for example, the components (SA) and (SR) in the sample are separated and become a constant velocity zone, and are transferred to the potential gradient detection type can. The potential curve at this time (D in the figure) is shown in FIG. In this way, the potential gradient of the terminal electrolyte is the largest (Po-P2) and the leading electrolyte is the smallest (P4-P), while the gradient of SB and sA (P2→P8) (P8 →p4) However, as the electrophoresis progresses, the terminal electrolyte T moves toward the detector (see A-D), resulting in electrophoresis TIIFJ'JdK (P in the figure). −、
-Po).

However, in the apparatus of the above embodiment, FIG.
As the terminal electrolyte moves, from a certain point (for example, when the voltage exceeds JKV), the electrode itself moves in the direction of the detector (42 (see Fig. 4, E)). voltage is cut, and the electrophoresis voltage becomes P
-Pl, and unnecessary voltage consumption can be avoided.

Further, no problem occurs with respect to the potential gradient of the components (SA) and (SR).

Although the above embodiments have been described as having means for moving the electrodes, basically any structure that allows the electrodes to be brought close to the potential gradient detector may be used. For example, as shown in FIG. Transfer tube (4) and terminal electrolyte tank (
2) Providing an electrophoresis tube moving block (goods) that slides the connection part with the electrophoresis tube in a liquid-tight state and brings it close to the electrophoresis double tube (with a transparent structure and the electrophoresis tube (4) as an electrode). The electrophoretic transfer unit (4) may be configured to control the electrophoretic voltage in the same manner as described above.As shown in FIG. 3 of L-1,
The electrode in the terminal electrode tank +21 is the main electrode (21a
) and an auxiliary electrode (21b) 7, an auxiliary electrode (21
+)) can be configured to be movable in the same way as the electrode (21) in FIG. 1, and configured so that a part of the voltage from the power source (5) can be applied thereto. Note that (51) is a control section that also includes a voltage supply section.

(f) Effects As described above, the isotachophoresis apparatus of the present invention requires a high voltage, and it is possible to lower the electrophoresis voltage. Therefore, the capacity and maximum voltage of the power supply for electrophoresis can be lowered, the isolation technique for the sample is simplified, there is no need to take special measures to improve the insulation of the system, and handling safety is improved. It is useful because it has various advantages such as improved performance.

[Brief explanation of the drawing]

FIG. 1 is a structural explanatory diagram showing one embodiment of the isokinetic electrophoresis device of the present invention, FIGS. 2 and 3 are structural explanatory diagrams showing other embodiments, respectively, and FIGS. 4A to 4E are FIG. 5 is a schematic diagram for explaining the state of use of the electrophoretic analyzer of the present invention, and a graph diagram for explaining the relationship between each zone and the potential during use. (iA,) (IB) (IC)... isotachophoresis device,
(21...Terminal electrolyte tank, (3)...Leading electrolyte tank, (4)...Migration tube, (6)...Constant current DC power supply,
H, H... electrode, (21a) - main electrode, (21b)
... Auxiliary electrode, (22! ... Electrode moving part, (22.
...Migration moving part, (exploitation... Patsukin, 41...
Moving block, (41)...Sample injection port, (4'4・
...Potential gradient detector, (4→...double tube, (51)
, (51)...control unit. Figure 3 Figure 4 Figure 5

Claims (1)

[Claims] 1. In an isotachophoresis apparatus in which a terminal electrolyte tank and a leading electrolyte tank are connected by a migration tube, and a sample injection port and a potential gradient detector are provided in the migration tube, "C" is used. an electrode moving means or an electrophoresis tube moving means capable of bringing an electrode set in a terminal electrolyte tank close to a potential gradient detector;
A constant-velocity electrophoresis device that is equipped with a voltage detector that monitors electrophoresis voltage during electrophoresis, and is configured to operate the moving means to prevent or reduce the electrophoresis voltage when the electrophoresis voltage exceeds a predetermined value. Electrophoresis device.