Two-dimensional capillary electrophoresis apparatus and use thereof

A capillary electrophoresis and capillary technology, which is applied in the preparation methods of peptides, organic chemistry, peptides, etc., can solve the problems that have not been constructed by 2D-CE, reduce the sample detection sensitivity, and the removal effect is not very complete, and achieve high peak capacity, Reduced replacement capacity requirements, easy-to-achieve effects

Inactive Publication Date: 2011-08-24
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, ampholytes absorb strongly at low UV wavelengths, reducing the detection sensitivity of samples
Although there are interfaces for removing ampholytes in the two-dimensional system constructed above, the removal effect is not very complete, which will affect the subsequent separation and detection
[Document 12: C. Yang, G. Zhu, L. Zhang, W. Zhang, Y. Zhang, Electrophoresis 2004, 25, 1729-1734.] by covalently bonding ampholytes to capillary monolithic columns Forming an immobilized pH gradient can effectively solve the above problems, but it has not been used in the construction of 2D-CE

Method used

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  • Two-dimensional capillary electrophoresis apparatus and use thereof
  • Two-dimensional capillary electrophoresis apparatus and use thereof
  • Two-dimensional capillary electrophoresis apparatus and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Such as figure 1 As shown, the first dimension adopts the M-IPG CIEF method, the positive buffer is 20mmol / L glutamic acid, and the negative buffer is 20mmol / L NaOH. The sample is filled with the entire M-IPG column, the overall column length is 20 cm (the lengths used in the present invention are 15, 20, 30 and 40 cm), and the inner diameter is 100 μm (can be 100, 200 and 250 μm). The second dimension adopts CZE to separate, and separation buffer is 20mmol / L glutamic acid, and capillary total length is 30cm (overlength adopted among the present invention is 20 and 30cm), and effective length is 20cm (overlength adopted among the present invention is 10 and 30cm). 20cm), the inner diameter is 50μm (can be 75μm).

[0042] The sample is a mixture of 7 standard proteins: trypsin inhibitor (trypsin inhibitor, pI 4.5), bovine serum albumin (BSA, pI 4.8), albumin (albumin, pI 4.9), β-lactoglobulin (β -lactoglobulin, pi 5.0), hemoglobin (Hemoglobin, pi 7.1), urease (urease, ...

Embodiment 2

[0047] The difference from Example 1 is that the sample used is protein extracted from milk, and the experimental operation is the same as that of Example 1.

[0048] Such as Figure 5 As shown, the separation of milk protein by one-dimensional M-IPG-CIEF is very poor, and the protein is not separated. Such as Figure 6 As shown, the separation results of proteins in milk have been significantly improved by using 2D-M-IPG-CIEF-CZE. Depend on Figure 7 It can be seen that the milk protein is better separated.

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Abstract

The invention relates to a two-dimensional capillary electrophoretic device, in particular to a novel two-dimensional capillary electrophoretic separation device and application thereof. The device can be used for the separation of a protein sample. The device comprises a 2D-CE interface. An immobilized pH gradient capillary monolithic column can be used as a first-dimension separation column of 2D-CE; the outlet end of the monolithic column is connected with one end of the 2D-CE interface through a connecting tube; and the sample inlet end of the monolithic column is arranged in an entrance tank containing alkaline buffer liquid or is connected with an injection pump. A capillary can be used as a second-dimension separation column of 2D-CE; the sample inlet end of the second-dimension separation column is connected with the other end of the 2D-CE interface through a connecting tube; and the outlet end of the second-dimension separation column is arranged in the tank containing the buffer liquid. The 2D-CE interface is arranged in a tank containing acidic buffer liquid. The device can remarkably improve the systematic peak capacity so as to meet the requirements of analyzing a complicated biosample.

Description

technical field [0001] The invention relates to a two-dimensional capillary electrophoresis device, in particular to a novel two-dimensional capillary electrophoresis (2D-CE) separation device and its application. Background technique [0002] Capillary electrophoresis (CE) has the advantages of high separation efficiency, fast analysis speed and less sample consumption. However, in the conventional CE method, due to the limitation of the applied voltage, the length of the separation column used is limited, which in turn affects the actual column efficiency and the possible peak capacity. In addition, due to the short optical path of the capillary, the detection sensitivity of CE is low. These disadvantages limit the application of CE in the analysis of complex biological samples. Therefore, it is of great significance to develop 2D-CEs with high peak capacity and high detection sensitivity by selecting orthogonal CE modes. [0003] At present, there have been many report...

Claims

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Application Information

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Patent Type & AuthorityPatents(China)
IPC IPC(8): C07K1/26
Inventor张丽华王婷婷朱贵杰孙良亮邓启良梁振张玉奎
OwnerDALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI