A Centrifugal Device for Enrichment and Separation of Phosphopeptides

Abstract

The invention discloses a centrifugal device for enrichment and separation of phosphopeptide. The centrifugal enrichment device includes an enrichment gun head, a separation gun head and a fraction collector. The enrichment gun head is filled with a solid phase extraction disk A and titanium dioxide filler, the solid phase extraction disk A is covered with an octalkyl film, the solid phase extraction disk A is close to a liquid outlet of the enrichment gun head, the separation gun head is filled with a solid phase extraction disk B and C18 reversed-phase filler, the solid phase extraction disk B is covered with an octadecyl film, and is close to a liquid outlet of the separation gun head; the enrichment gun head can be disposed in the separation gun head; and the enrichment gun head and the separation gun head both can cooperate with the fraction collector. The centrifugal device provided by the invention achieves organic combination of phosphopeptide enrichment and reversed-phase separation, effectively simplifies the experimental steps, reduces the sample loss, is very suitable for phosphopeptide analysis of large-scale trace clinical samples, and has good application potential in enrichment study of proteome posttranslational modification.

Description

technical field [0001] The invention relates to a centrifugal device for phosphopeptide enrichment and separation, which belongs to the field of analytical chemistry. Background technique [0002] Phosphorylation is one of the most common post-translational modifications of proteins, which refers to the addition of covalently bound phosphate groups on amino acid residues of substrate proteins catalyzed by protein kinases. The reverse reaction is the dephosphorylation process catalyzed by protein phosphatases. Protein kinases and phosphatases work independently and in balance to regulate protein function. Studies have shown that about 30% of the proteins encoded by genes will be phosphorylated. The reversible process of protein phosphorylation and dephosphorylation regulates almost all processes of life activities, including cell proliferation, development and differentiation, molecular recognition and signal transduction, nerve activity, muscle contraction, metabolism, tum...

AI Technical Summary

Problems solved by technology

However, there are still some problems: (1) the initial amount of sample required is large, (2) the experimental steps are cumbersome, which is extremely time-consuming and laborious, (3) the throughput of sample analysis is limited, (4) the reproducibility of enrichment and identification results is relatively low. Difference

However, the currently widely used reversed-phase chromatography column peptide separation mode has a large sample loss, and needs to be used with liquid chromatography instruments, and the sample processing throughput is limited.

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