Phosphorylated peptide adsorption enrichment method, additive and application

Abstract

The invention discloses a phosphorylated peptide adsorption enrichment method comprising (1) a non specific adsorption inhibitor and part of a to-be-tested sample are added a sample loading buffer, and an enrichment filter column is used for phosphorylated adsorption enrichment; (2) enrichment products are collected, the abundance of 20 kinds of amino acids in non phosphorylated peptide can be calculated as first abundance, the abundance of the non phosphorylated peptide in the 20 kinds of amino acids can be calculated as second abundance, at least one amino acid with higher ratio of first abundance to second abundance can be selected as an additive; (3) the sample loading buffer added with the additive is used for phosphorylated adsorption enrichment of the to-be-tested sample. By use of the competitive adsorption effects of amino acids and peptides, adsorption of the non phosphorylated peptide can be reduced, adsorption selectivity of the phosphorylated peptide enrichment can be increased, and the method lays the foundation for improving the accuracy and stability of phosphorylation detection.

Description

technical field [0001] This application relates to the field of enrichment and purification of proteins and polypeptides, in particular to an adsorption enrichment method for phosphorylated peptides, an additive for phosphorylation enrichment, a preparation method and application of the additive. Background technique [0002] In biological cells, proteins are the main function performers. Due to functional needs, many proteins will covalently bind various non-peptide groups on the amino acid chain during or after translation to form post-translational modifications. Phosphorylation of proteins is one of the most important post-translational modifications of proteins, and reversible phosphorylation controls cellular activities such as cell proliferation, cell division, intercellular information transmission, and gene expression. However, in protein samples, the content of phosphorylated proteins and phosphorylated peptides is very low, and there are interferences from various...

AI Technical Summary

Problems solved by technology

However, the effect of existing non-specific inhibitors in phosphorylated peptide enrichment is not good, especially in the currently commonly used titanium dioxide enrichment method, the enrichment selectivity of existing non-specific inhibitors is generally 60% -70% or so; that is, there are still 30%-40% or more non-phosphorylated peptides enriched, which seriously affects the accuracy and stability of phosphorylated peptide detection

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