Deep proteomics analysis method for trace sample and application

By introducing high-field asymmetric waveform ion mobility spectroscopy (FAIMS) into the liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, the sample processing and mass spectrometry detection parameters were optimized, and the problems of insufficient coverage and poor quantitative stability in the proteomic analysis of micro samples were solved, thereby achieving high-deep and high-sensitivity proteomic analysis.

CN119688889AActive Publication Date: 2025-03-25THE EYE HOSPITAL OF WENZHOU MEDICAL UNIVERSITY
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Patent Information

Application Number
CN202510193983.3
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-02-21
Publication Date
2025-03-25
Estimated Expiration
2045-02-21

AI Technical Summary

Technical Problem

When performing proteomic analysis in trace samples (such as retinal pigment epithelium/choroid complex), the resolution is low, the coverage is insufficient, the dynamic range is limited, the detection rate of low abundance proteins is low, and the quantitative repeatability and background noise suppression are insufficient, which affects the accurate identification of disease-related proteins.

Method used

Deep proteomic analysis of microscope-tandem mass spectrometry (LC-MS/MS) was performed on microscope-induced proteomics using high-field asymmetric waveform ion mobility spectrometry (FAIMS). The method includes sample processing, liquid chromatography separation, FAIMS separation and mass spectrometry detection, and improves separation efficiency and dynamic range by optimizing parameters such as carrier gas flow rate, compensation voltage and mass spectrometry scanning conditions.

Benefits of technology

High-deep and high-sensitivity proteomic analysis of trace samples is achieved, the number of protein identification, dynamic range and quantitative repeatability are improved, the proportion of missing values ​​is reduced, and low-abundance proteins and cell type markers can be detected, which helps in the study of disease mechanisms.

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Abstract

The invention relates to a deep proteomic analysis method aiming at a trace sample and application thereof, which are used for high-depth and high-sensitivity proteomic analysis of a retinal pigment epithelium / choroidal complex, are also suitable for proteomic deep testing of other trace samples, have wide applicability and flexibility, and can be widely applied to industrial production. According to the method, LC-FAIMS-MS / MS parameters are systematically optimized, the technical bottlenecks of low coverage of trace tissue proteome and poor quantitative stability are overcome, a high-sensitivity solution is provided for disease mechanism research and biomarker discovery, the protein identification number is increased by 28%, the dynamic range is expanded to 6 orders of magnitude, quantitative repeatability is improved, and the method is suitable for clinical application. The missing value proportion is reduced to 6%; aMD-related low-abundance proteins and 10 cell type markers can be detected, and disease mechanism research is assisted.
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