Rapid treatment method for phosphoproteome sample

Abstract

The invention relates to a rapid treatment method for a phosphoproteome sample and develops a novel sample treatment method integrating cell splitting decomposition and protein enzymolysis by using the advantage that protein rapid enzymolysis can be promoted through high-concentration trypsin, and application of the novel sample treatment method in analysis of phosphoproteomics. Benefiting from the high-concentration trypsin, a deep mechanism of the cell splitting decomposition and the protein enzymolysis is promoted; a cell sample can finish rapid conversion of a peptide fragment mixture in one step; and in a phosphoeptide enriching process, a non-phosphorylated peptide fragment and other substances with incompatible mass spectrums can be removed. The rapid treatment method can realize the rapid conversion from cells to the peptide fragment by 25 minutes; in a contrast normal group, the simplest method also needs at least 16 hours; and a relatively good mass spectrum identification coverage rate of the contrast group is obtained through analyzing a HeLa actual cell sample, and the accuracy and the high efficiency of the sample pre-treatment method are proved.

Description

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AI Technical Summary

Benefits of technology

In this new technology described by our researchers, it suggests quickly analyzing large amounts (>10 grams) of proteins with specific properties called Phosphoprotein or Protease K). These techniques are useful because they allow us to study how these molecules work together more efficiently during various biological processes like metabolism, signal transduction pathways, gene regulation, etc., which could lead to important applications ranging from drug development to diagnositic purposes.

Problems solved by technology

This patents describes various technical solutions aim at improving the speed and accuracy of analyzing large amounts of data collected from experiments involving studying specific aspects of cells or tissues during their physiological processes like growth, gene expression dynamics, histochemistry, molecular imaging, and drug discovery. Current techniques involve multiple stages of liquid phase separation followed by enzymic activity measurement after each stage. These techniques require expensive equipment and consume considerable amount of time. Additionally, they may result in reduced sensitivity due to poor substrate selectivity towards lysochlorophyll breakdown.

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