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

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...

AI Technical Summary

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

Images