Untargeted Proteomics Sequencing

Untargeted proteomics sequencing aims to comprehensively analyze all proteins in a biological sample, rather than focusing on specific proteins, and contributes to studying the complexity of the proteome and discovering new protein biomarkers. Protein sequencing is the process of determining the amino acid sequence of a protein. Furthermore, Untargeted protein sequencing, also known as shotgun proteomics, aimed at analyzing all proteins in a biological sample, rather than selectively determining specific target proteins.

 

Features of Untargeted Proteomics Sequencing

1. Wide Coverage

It attempts to identify all detectable proteins in the sample, not limited to a specific group of proteins.

 

2. Discovery-Oriented

This method is suitable for exploratory research, where the research goal is not to validate a specific hypothesis, but to discover new patterns, connections, or protein functions.

 

Workflow

1. Sample Preparation

Proteins are extracted from the biological sample and appropriately treated, such as protein degradation, reduction, and alkylation.

 

2. Liquid Chromatography Mass Spectrometry Analysis

Liquid chromatography mass spectrometry (LC-MS/MS) is used to separate and spectrometrically measure the protein sample. This can fragment the proteins in the sample into many pieces that can be used for identification and quantification.

 

3. Data Analysis

The mass spectrometry data is compared with known protein sequences in the database to identify the proteins present in the sample. Moreover, methods for analyzing unknown proteins can also be used to identify new proteins.

 

4. Quantitative Analysis

The intensity of the mass spectrometry signal is used to quantify each identified protein, to understand their relative abundance in the sample.

 

5. Bioinformatics Analysis

The identified and quantified proteins are associated with relevant information such as biological function, pathways, and metabolic networks, to gain more biological insights about the sample.

 

Application Fields

1. Discovery of Biomarkers

In fields such as disease diagnosis and monitoring of drug treatment responses, untargeted proteomics can be used to discover new biomarkers.

 

2. Disease Mechanism Research

Comparing the proteomes of tissues or cells in healthy and disease states can help understand the molecular mechanisms of diseases.

 

3. Drug Mechanism of Action

Exploring the effects of drug treatment on the cellular proteome can reveal the molecular targets and mechanisms of action of drugs.

 

4. Systems Biology Research

In systems biology, untargeted proteomics helps build protein networks of cells and tissues and understand complex biological processes.