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Two-dimensional conventional column array type chromatographic separation system and method for removing high-abundance proteins

A chromatographic separation and protein technology, which is applied in the field of two-dimensional conventional column array chromatographic separation systems, can solve the problems of low sample load, unfavorable enrichment and separation analysis of high-abundance proteins and low-abundance proteins, and achieves improved detection sensitivity, The effect of improving separation throughput and shortening sample analysis time

Inactive Publication Date: 2015-05-13
FUDAN UNIV
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Problems solved by technology

At present, two-dimensional array capillary liquid chromatography-matrix-assisted laser desorption ionization tandem mass spectrometry (MALDI-MS-MS) system has been used for the research of proteomics analysis, but for complex samples, the capillary array is difficult due to the The load is relatively small, which is unfavorable for the removal of high-abundance proteins and the enrichment and separation of low-abundance proteins

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  • Two-dimensional conventional column array type chromatographic separation system and method for removing high-abundance proteins
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  • Two-dimensional conventional column array type chromatographic separation system and method for removing high-abundance proteins

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Embodiment Construction

[0025] 1. The establishment of a two-dimensional conventional column array chromatographic separation system for the removal of high-abundance proteins.

[0026] 1. Design and manufacture of multi-channel shunt 7:

[0027] A stainless steel block (with internal thread) with a diameter of 20mm and a discontinuous circular groove engraved at the position of 10mm in diameter, with 10 micro-circular holes with a diameter of 0.5mm evenly distributed in the circular groove on the top, a diameter of 12mm, thickness 3mm polyimide gasket, a 12mm diameter, 1.5mm thick stainless steel gasket, a 20mm diameter stainless steel gasket with micro holes drilled and a set of polyimide gaskets for fastening to make the system tight and sealed bolts.

[0028]When in use, 10 capillaries of the same specification (20 cm × 320 mm i.d. 400 mm o.d.) can be fixed into the micropores of the stainless steel block, and tightened with bolts to make the polyimide gasket deform, so that the seal at the inte...

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Abstract

The invention belongs to the technical field of proteomics, and in particular relates to a two-dimensional conventional column array type chromatographic separation system for removing high-abundance proteins and a separation method applying the system. According to the invention, only one multi-element liquid phase pump, one ten-way valve and two six-way valves are needed for connection and on-line two-dimensional switching; and the automation degree and the integration degree of an instrument are improved. Fractions obtained by performing first-dimensional IE separation on a protein sample are directly and sequentially switched through the ten-way valve and transferred to a column head of a multi-channel second-dimensional conventional reversed-phase column for enrichment; the reversed-phase column directly performs on-line desalting and gradient dilution on the protein sample; by two-dimensional conventional chromatographic separation, the high-abundance proteins can be precisely positioned on a two-dimensional chromatogram; and therefore, the accurate removal aim is fulfilled. By the method, large-volume sample injection, concentration desalting, array type separation and positioning and removal of the high-abundance proteins can be finished; and the two-dimensional conventional column array type chromatographic separation system is very suitable for positioning, removal and high-flux analysis for the high-abundance proteins required by research on proteomics.

Description

technical field [0001] The invention belongs to the technical field of proteomics, and in particular relates to a two-dimensional conventional column array chromatographic separation system for removing high-abundance proteins, and a separation method using the system. Background technique [0002] With the completion of human genome sequencing, the focus of life science research has also shifted to the study of proteomics. Its main problems focus on the large-scale identification of unknown proteins and the determination of low-abundance proteins. In recent years, high performance liquid chromatography (HPLC) has played an increasingly important role in proteomics as an efficient separation method. Compared with the limited separation ability of one-dimensional separation, multi-dimensional separation technology has larger peak capacity, fast separation speed, good reproducibility, and high degree of automation. Therefore, the development of multi-dimensional orthogonal li...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01D15/08G01N1/34
Inventor 张祥民高明霞洪广峰黄志
Owner FUDAN UNIV
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