Method and apparatus for controlling electrovadose in capillary isoelectrofocusing electrophoresis

An isoelectric focusing electrophoresis and capillary technology, which can be applied to measurement devices, separation methods, chemical instruments and methods, etc., can solve the problems of flow pattern change and separation effect deterioration, and achieve no change in flow pattern, improve focusing effect, separation good effect

Inactive Publication Date: 2008-03-26
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although a pressure-driven scheme can be used to make the sample flow out of the column, the pressure-driven result will change the flow pattern and deteriorate the separation effect

Method used

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  • Method and apparatus for controlling electrovadose in capillary isoelectrofocusing electrophoresis
  • Method and apparatus for controlling electrovadose in capillary isoelectrofocusing electrophoresis
  • Method and apparatus for controlling electrovadose in capillary isoelectrofocusing electrophoresis

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Experimental program
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Effect test

Embodiment 1

[0035] Referring to Figure 1, the present invention provides a device for controlling electroosmotic flow in capillary isoelectric focusing electrophoresis, which is composed of a capillary separation column 1, a heating device 2, a freezing device 3, a detector 4 and a power supply. The front end of the capillary separation column 1 is The sample inlet 6, the rear end is the sample outlet 7, and are respectively connected to the power supply (not shown in the figure). The front ends of the heating device 2 and the freezing device 3 are respectively provided with gas injection holes 21 and 31, and are arranged in the capillary separation On both sides of the appropriate position of the column 1, that is, one end of the sample outlet 7 of the capillary separation column, the detector 4 is installed on the capillary separation column 1 between the heating device 2 and the sample outlet 7.

[0036] According to the principle that the fluid will condense into a solid at low temperatur...

Embodiment 2

[0042] Please refer to Figure 2. The present invention provides another device for controlling electroosmotic flow in capillary isoelectric focusing electrophoresis. It consists of two capillary separation columns 1 and 5, a heating device 2, a freezing device 3, a detector 4 and a power supply (not shown in the figure). Given) composition, the front end of the capillary separation column 1 is the injection port 6, the back end is connected to the interface 8, and the interface 8 is connected to the power supply, the flow path 9 and the capillary column 5 at the same time, and the end of the capillary column 5 is the sample outlet 7. Electrodes B and C are inserted into the sample inlet 6 and the sample outlet 7 respectively. Electrode A is inserted at the upper end of the interface 8, and its lower end is a flow path 9 with a shut-off valve 91, which is used to empty part of the fluid in the tube, and can also be used for the separation of multi-dimensional isoelectric focusing; ...

Embodiment 3

[0045] Example 3: Electro-osmotic flow changes

[0046] Experimental conditions: (using the separation device described in Example 1)

[0047] Capillary (Hebei Yongnian Ruiyang Chromatography Device Co., Ltd., inner diameter: 75μm, outer diameter: 375μm), the inner surface is inertly treated; carrier ampholyte Pharmalyte (pH3.0-5.0), N, N, N′, N′-tetramethylethylenediamine (TEMED, 99%). TriSep of Unitech TM -2000GV type capillary electrophoresis instrument, including a tunable wavelength ultraviolet-visible detector, a continuously adjustable high-voltage DC power supply.

[0048] Working voltage: 5kv

[0049] Liquid nitrogen freezing: freezing time 2s

[0050] Hot air heating: heating time 2s

[0051] Referring to Figure 3, it can be seen that the electroosmotic flow remains at a stable value during the freezing process, and after heating, the electroosmotic flow returns to the initial state.

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Abstract

The present invention discloses a method for controlling electroosmotic flow in capillary isoelectric focusing electrophoresis and its equipment. It is characterized by that it adopts freeze method to freeze local fluid in the capillary to make focusing of fluid, then adopts the method of heating capillary to defreeze the solid mobile phase in the capillary and restore the transmission of fluid in the capilalry, so that the flowing-out of the sample can be batch implemented, and the focusing effect of the capillary isoelectric focusing electrophoresis can be greatly raised, and the flowing form in the column is wedge-shaped, and its separation effect is good. On the proper position of capillary separation column a heating device and a freezing device are set.

Description

Technical field [0001] The invention provides a separation method and device used in capillary isoelectric focusing electrophoresis. Background technique [0002] Capillary isoelectric focusing electrophoresis method used to separate bisexual samples has advantages that other separation modes cannot match. In this separation mode, a capillary with a treated inner surface is generally used. On the one hand, in order to reduce the adsorption of proteins and other samples on the tube wall, on the other hand, reduce the number of silanol active sites on the tube wall as much as possible to reduce the electroosmotic flow. . Electroosmotic flow is one of the most important factors affecting the focusing of components in the isoelectric focusing separation process. No matter what pipe wall treatment method is used, it is impossible to reduce the electroosmotic flow to zero, thus limiting the focusing ability of the separation process. [0003] During the focusing process, the presence o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447C07K1/28B01D57/02
Inventor 张玉奎张维冰张庆合张凌怡杨春
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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