Non-targeted proteomics aims to comprehensively analyze all proteins in a biological sample, rather than focusing on specific proteins, helping to study the complexity of the proteome and discover new protein markers. Protein sequencing is the process of determining the amino acid sequence of a protein. It is an important technique in biological research to understand the structure and function of proteins, as well as to study the biological processes associated with proteins. Non-targeted protein sequencing is a comprehensive proteomics approach that aims to analyze all proteins in a biological sample, rather than selectively determining a specific protein of interest. This method is also known as shotgun proteomics.
Features of non-target proteomic sequencing
1.Broad coverage: Try to identify all detectable proteins in the sample, not just a specific group of proteins. 2.Discovery-oriented: This approach is suitable for exploratory research where the goal is not to test a specific hypothesis, but to discover new patterns, connections, or protein functions. The general steps of non-target proteomic sequencing include:
1. Sample preparation:
Proteins are extracted from biological samples and treated appropriately, such as protein degradation, reduction, and alkylation.
2. Liquid chromatography mass spectrometry analysis
Protein samples were separated and determined by mass spectrometry using liquid chromatography mass spectrometry (LC-MS). This allows the protein in the sample to be divided into many fragments that can be used for identification and quantification.
Figure 1Untargeted proteomics.
3. Data analysis
Align mass spectrometry data with known protein sequences in the database to identify the proteins present in the sample. In addition, analytical methods for unknown proteins can also be used to identify new proteins.
4. Quantitative analysis
Each identified protein is quantified by the intensity of the mass spectrometry signal to understand their relative abundance in the sample.
5. Bioinformatics analysis
Correlate identified and quantified proteins with relevant information about biological functions, pathways, and metabolic networks to gain additional biological insights into your samples.
Fields of application
1.Biomarker discovery: Non-target proteomics can be used to discover new biomarkers in areas such as disease diagnostics and drug response monitoring. 2.Disease mechanism studies: By comparing the proteome of tissues or cells in healthy and disease states, it can help to understand the molecular mechanisms of disease. 3.Mechanism of action: To explore the effects of drug treatment on the cellular proteome, and to reveal the molecular targets and mechanisms of action of drugs. 4.Systems biology research: In systems biology, non-target proteomics helps to build protein networks in cells and tissues to understand complex biological processes.