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Column-switching circulating volume exclusion chromatography system for biomacromolecule separation and purification

A technology of size exclusion chromatography and biological macromolecules, applied in the field of protein separation and purification systems, can solve problems such as overlapping chromatographic peaks, and achieve the effects of improved resolution, high resolution and simple design

Inactive Publication Date: 2010-01-06
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

However, as the separation time increases, the spectral peaks broaden, and the separated chromatographic peaks (proteins, nucleic acids, etc. with a wide molecular weight range) may overlap after multiple cycles of separation

Method used

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  • Column-switching circulating volume exclusion chromatography system for biomacromolecule separation and purification
  • Column-switching circulating volume exclusion chromatography system for biomacromolecule separation and purification
  • Column-switching circulating volume exclusion chromatography system for biomacromolecule separation and purification

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Such as figure 1 As shown, a column switching cycle size exclusion chromatography system for the separation and purification of biomacromolecules, the device consists of a tee joint 1, a pump 2, an injection valve 3, a size exclusion chromatography column A4, a switching valve A5, a volume The exclusion chromatographic column B6, the detector 7, and the switching valve B8 are composed, and they are connected in the form of a closed loop through pipelines.

[0028] One end of the size exclusion chromatographic column A4 is connected to the injection valve 3, and the other end is connected to the size exclusion chromatographic column B6 and the detector 7 respectively through a switching valve A5; The through joint 1 is respectively connected with the external mobile phase source and the outlet of the detector 7; a switching valve B8 is arranged on the connecting pipeline between the three-way joint 1 and the detector 7, and the detector 7 is connected with the collection...

Embodiment 2

[0034] Such as image 3 As shown, the device consists of three-way connector 1, pump 2, injection valve 3, size exclusion chromatography column A4, switching valve A5, size exclusion chromatography column B6, ultraviolet detector 7, valve B8, pump B17, and pump C21 , pump D22, switching valve C9, C8 trapping column A13, C8 trapping column B14, reverse-phase chromatographic column 15, and ultraviolet detector B16.

[0035] The difference from Example 1

[0036] The switching valve B8 is a ten-way valve, the interface connected to the detector 7 is ①, and the remaining interfaces clockwise are ②③④⑤⑥⑦⑧⑨⑩, the interface ① is connected to the detector 7, the interface ⑩ is connected to an interface of the three-way joint 1, The interface ③ is connected with the external mobile phase D18 through the pump B17, and the interface ② is connected with a switching valve C9;

[0037] Switching valve C9 is a ten-way valve, the interface connected to switching valve B8 is ①, and the remain...

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Abstract

The invention relates to a column-switching circulating volume exclusion chromatography system for biomacromolecule separation and purification, which comprises a liquid chromatography pump, a sampling valve, a three-way or four-way connector, a switching valve A, a switching valve B, a volume exclusion column A, a volume exclusion column B and a detector. The pump, the sampling valve, the volume exclusion column A, the switching valve A, the volume exclusion column B, the detector, the switching valve B and the three-way or four-way connector are connected with one another in turn through pipelines, the number of the biomacromolecules getting into a circulation and separation system can be controlled by adjusting the switching frequency or mode of the valve A, and the separation and the collection of the biomacromolecule components can be controlled by adjusting the switching of the valve B. Compared with the conventional volume exclusion chromatography, the system has the characteristics of strong separating power, high resolution and the like, and can be applied to the separation and the purification of biomacromolecule samples. Simultaneously, the system also has good utility value in constructing a continuous biomacromolecule multi-dimensional separation platform.

Description

technical field [0001] The invention relates to a protein separation and purification system, in particular to a column switching cycle size exclusion chromatography separation system. Background technique [0002] In the separation of biological macromolecules, ion exchange chromatography and size exclusion chromatography are the two most commonly used separation techniques. Size exclusion chromatography is often used in the separation and purification of biomolecules due to its mild separation conditions and easy preservation of the activity of biomolecules. Compared with other separation modes, SEC is considered to be a low-resolution liquid chromatography method, thus limiting its application in the separation of biomacromolecules. In order to improve the resolution, longer columns or columns packed with finer particles are usually used, but the column pressure is too high to make this method difficult to put into practice. Cyclic size exclusion chromatography is a liq...

Claims

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Application Information

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IPC IPC(8): C07K1/16C12N15/10B01D15/34C07H1/06
Inventor 张丽华袁辉明梁振张玉奎
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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