Whole protein online fractionation pre-separation and separation analysis method and its liquid chromatography system

Abstract

The invention provides an on-line pre-separating and separating analysis method for a whole protein and a liquid chromatogram system thereof. By utilizing orderly proportionately increased characteristic of hydrophobicity of Cn capture column, proteins in a protein mixture are orderly gradingly captured from large to small to implement pre-separation; and the proteins on the capture column are orderly matched by relative Cn analytical columns to implement secondary separation, and the analytical column is used together with the mass spectrum for analysis. The orderly proportionately increased capture columns Cn (n=1-18 or partial combination thereof, such as 1, 2, 4, 8 and 18) are used for on-line pre-separating samples, which greatly raises a pre-separation speed, sensitivity and resolution; and the separation columns and the capture column are pairwise matched (such as C4 capture and C4 separation column are matched), which have consistent hydrophobicity, thereby implementing maximization of separation resolution.

Description

technical field [0001] The invention relates to a technology and method for on-line fractionation, pre-separation and separation analysis of whole proteins. Background technique [0002] In today's post-genome era, the entire genome sequences of many species have been or are being determined, which makes comprehensive and in-depth research on the proteome possible. Since most of the biological functions are ultimately transmitted and realized through proteins, the research on proteins will promote the development of biology and medicine more directly and rapidly on the basis of gene research. The technology based on high performance liquid chromatography-mass spectrometry can quickly and sensitively analyze and identify proteins and their mixtures, and is the frontier and preferred research method for protein research. In the past ten years, mass spectrometry-based protein analysis and identification technology has been developed rapidly, and it has been able to perform com...

AI Technical Summary

Problems solved by technology

Since the sample loss in the desalting process is very serious and time-consuming, the analysis speed is slow and the sensitivity is low when ion exchange chromatography is used for pre-separation

The eluent of size exclusion chromatography is compatible with liquid chromatography-mass spectrometry analysis, can be used online, and the analysis speed is fast, but its resolution is often very low

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