JPH06198137A - Recovery part for object to be inspected of fractional collecting electrophoresis device and recovery method for the same - Google Patents

Abstract

PURPOSE:To provide a fractional collecting device and recovery where there remains no recovered liquid in a recovery chamber and components of an object to be inspected are recovered at low dilution. CONSTITUTION:A recovery chamber 11 is constituted of a hollow recovery chamber forming body 9 into which upper and lower glass tubes 5, 7 are inserted from above and below respectively. The inserted end faces of the upper and lower glass tubes 5, 7 are worked together with a gel-like body so as to form convex faces. The outer circumferential edges of the convex faces are made to substantially coincide with the levels of ports 11a, 11b, 11c, 11d from the outer surface of the hollow recovery chamber forming body 9 to the hollow part of it so that there may remain no recovered liquid in the recovery chamber. Further in a fractional collecting method, the recovery chamber is filled with the recovered liquid and one component separated and leaked is dissolved in recovery liquid C and the recovered liquid containing the component is taken out from a recovered liquid take-out port by replacing it with air and this operation is repeated every time each component is leaked to lower the.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to the shape of a test substance recovery section in a preparative electrophoresis apparatus and a method for recovering the test substance.

[0002]

Background of the Invention What is a preparative electrophoresis apparatus? Electrophoresis separates a test substance component on the upper end of the gel-like support into a layer in the gel-like support, and further fractionates the gel-like support. Is a device for continuously flowing an electric current to leak the components separated in layers from the lower end of the support. This operation of taking out the components is called collection, and the collection of each component can be achieved by repeating the collection for each component.

The major part of the preparative electrophoresis apparatus is a separation support for separating the contained test substance, a recovery chamber for recovering components leaking from the lower end of the separation support, and a separation support. It consists of a lower gel-like body that faces the bottom and becomes the bottom of the recovery chamber. The recovery chamber is formed between the lower end of the separation support and the upper end of the lower gel-like body. Both the separating support and the lower gel-like body are usually produced in a cylindrical glass tube. Two tubes, one outlet for the recovered solution and one inlet for the recovered solution, are connected to the recovery chamber.

When the preparative electrophoresis apparatus is energized, the test substance moves to the recovery chamber while being separated into each component in the separation support. The recovery solution is always sent to the recovery chamber, and the component of the test substance leaked from the lower end of the separation support is immediately transferred to the recovery solution flow and carried outside the preparative electrophoresis apparatus. The collected solution containing the components of the test substance carried to the outside of the preparative electrophoresis apparatus is collected in a fraction collector for each component.

[0005]

2. Description of the Related Art In a conventional apparatus, it takes a long time for the recovered solution to be constantly sent at a constant flow rate so that one component as a whole leaks out, so that the amount of recovered liquid per component becomes large and a separation support is required. Compared with the concentration of the test substance in the body, it is significantly diluted and recovered. Further, if the flow of the recovery liquid for preventing this dilution is interrupted, air remains in the recovery chamber or the recovery liquid for the previous recovery remains, which is inconvenient. Furthermore, bubbles generated due to the rise in liquid temperature due to energization may remain in the recovery chamber. In order to eliminate these drawbacks, there are some devices in which the electrophoresis device is made horizontal and the collection chamber is arranged vertically, but this is not applicable when the test substance is in the form of a solution. Since the moving direction of the test substance is orthogonal to the direction in which gravity is applied, the test substance diffuses in two directions and impairs separation.

[0006]

Therefore, the present invention has improved the shape of the recovery chamber so as to avoid the above-mentioned drawbacks even though the recovery liquid is not continuously fed.

[0007]

In the preparative electrophoresis apparatus according to the present invention, the separating gel support, the lower gel body and the collecting chamber are all completely immersed in the preparative buffer. It is possible to maintain a constant temperature by controlling the temperature of the migration buffer.

If the recovery liquid is continuously sent as it is, the recovery can be performed as in the conventional case, but the problem of dilution cannot be solved. For the problem of dilution, fill the recovery chamber with the recovery liquid, stop supplying liquid until one component leaks while energizing,
After that, the problem can be solved by sending air into the recovery chamber and taking out the solution in the recovery chamber. The recovery chamber filled with air is filled with the recovery liquid again, and the leakage of the next component is waited for. If air remains in the collection chamber, it will hinder energization and prevent normal separation. To fill the recovered liquid without leaving air, provide an inlet for the recovered liquid at the lower position of the recovery chamber and an outlet at the highest position.
The lower end of the separation support at the upper part of the recovery chamber is made to be convex downward. This convex shape includes a dome shape, a cone shape, a trapezoidal shape, and the like. When the lower end of the separating support itself is flat, the edge of the glass tube may be scraped off so that the entire lower end has a convex shape.

To take out the solution in the recovery chamber without leaving any residue, the upper end of the lower gel-like body is made convex so that the liquid does not remain attached to the upper end of the lower gel-like body in the lower part of the recovery chamber. Like the tube, the processing of the upper edge of the lower gel glass tube should be gradually deeper toward the solution outlet,
By setting the outlet of the solution to the bottom of the recovery chamber, it is possible to take out even a small amount of liquid.

[0010]

FIG. 1 shows a longitudinal section of a preparative electrophoresis apparatus according to the present invention. The electrophoresis tank is composed of an upper electrophoresis tank 1 and a lower electrophoresis tank 3, each filled with electrolyte solutions A and B. The negative electrode 2 and the positive electrode 4 are immersed and installed, respectively. The recovery chamber forming body 9 attached to the bottom of the migration tank 1 has a port 11a,
11b, and ports 11c, 1 facing each other in diameter.
1d is provided.

The separating support 6 is formed in the upper glass tube 5, and the inclined surface 5a is chamfered around the entire lower edge of the glass tube 5, and the outer periphery of the inclined surface 5a and the ports 11a and 11c are formed. The glass tube 5 is inserted into the forming body 9 so that the level of the uppermost portion coincides with that of the uppermost portion, and an O-ring 10 fixes and seals between them. Similarly, a lower support 8 is formed in the lower glass tube 7 so that the outer circumference of the maximum inclined surface 7a having the maximum slope in the chamfering of the upper edge of the glass tube 7 and the lower portion of the port 11b are formed. The glass tube 7 is inserted into the forming body 9 so that the levels coincide with each other, and an O-ring 10 'is used to fix and seal between them.
As described above, the recovery chamber 11 is formed between both gel-like bodies.

The pump 12 sucks the recovery liquid C and fills the recovery chamber 11 with the recovery liquid C via the recovery liquid inflow port 11d and the outflow port 11c. The excessively sucked liquid is stored in the container 13, and after the complete filling, the pump 12 is stopped.
At this time, the air valve 14 is closed and the pump 15 is stopped.

At the time of electrophoresis, the test substance 19 is placed on the upper end of the separation support 6 and electricity is applied. After the component of the test substance moves downward while being separated, and one component of the test substance leaks to the recovery chamber 11, the energization is stopped, the air valve 14 is opened, and suction by the pump 15 is started. Air supply port
Air is sent into the recovery chamber 11 from the port 11a, and the recovery liquid containing the test substance component passes through the extraction port 11b,
Collected in a test tube of the dispenser 16. The detector 17 and the recorder 19 detect and record a specific component. After the collection, the empty collection chamber 11 is filled with the collection liquid C again, and the same operation is performed. After filling the collection chamber 11, energization is started. In this way, each of the components contained in the test substance 19 is tested in a test tube by repeating the cycle of filling the recovery liquid for each leaking component-starting energization-leakage of test substance components-stopping energization-collection and sampling. It can be collected separately.

[0014]

The structure of the recovery chamber according to the present invention has no recovery liquid remaining in the recovery chamber, and the recovery method according to the present invention allows the recovery liquid to flow every time a component leaks. Since it does not flow through, the analyte components can be recovered at low dilution.

[Brief description of drawings]

FIG. 1 is a vertical cross-sectional view of a preparative electrophoresis apparatus of the present invention.

[Explanation of symbols]

 1: Upper electrophoresis tank 3: Lower electrophoresis tank A, B: Electrolyte solution 5: Upper glass tube 5a: Inclined surface 6: Gel support for separation 7: Lower glass tube 7a: Maximum inclined surface 8: Lower gel shape Body 9: Recovery chamber forming body 11a: Air supply port 11b: Extraction port 11c, 11d: Recovery liquid inflow / outflow port 11: Recovery chamber C: Recovery liquid 12, 15: Pump 14: Air valve 16 : Separator 19: Recorder

Claims (3)

[Claims] 1. An end surface of an upper and a lower glass tube in which a gel support for separation and a lower gel are respectively made to face each other are vertically opposed to each other, and a recovery chamber for a test substance component separated by electrophoresis is provided between them. In the preparative electrophoresis apparatus thus formed, the recovery chamber comprises a hollow recovery chamber forming body in which upper and lower glass tubes are inserted from above and below, respectively, and the end faces into which the upper and lower glass tubes are inserted are gel-like. It is processed so as to form a convex surface with the body, and the outer peripheral edge of the convex surface is made substantially coincident with the level of the port from the outer surface of the recovery chamber forming body to its hollow portion. Electrophoretic device. 2. A pair of ports for inflow and outflow of the recovered liquid.
Port, an air supply port, and a recovery liquid removal port containing the test substance. The recovery liquid inflow port and the recovery liquid removal port are convex surfaces on the end face of the lower glass tube. The preparative electrophoresis apparatus according to claim 1, wherein the level of the recovery liquid take-out port is slightly lower than the level of the recovery liquid inflow port so that the level thereof is matched with the outer peripheral edge. 3. A method for collecting each component contained in a test substance using the electrophoresis apparatus for separation according to claim 2, wherein the recovery chamber is filled with a recovery liquid, and the separation is performed. Each component is prepared by dissolving one of the components separated and leaking from the lower end surface of the gel-like support for use in the recovered liquid, and replacing the recovered liquid containing this component with air to take out the recovered liquid from the recovery liquid extraction port. A method of recovering a test substance component, which is characterized in that it is repeated every time a substance leaks.