Single-Molecule Proteomics

Single-molecule proteomics (SMP) is an advanced proteomics technology designed to detect, quantify, and analyze proteins at the single-molecule level. Unlike traditional proteomics, which relies on large sample sizes, single-molecule proteomics enables the study of protein diversity, heterogeneity, and post-translational modifications (PTMs) using minimal sample amounts or even single cells. This groundbreaking approach provides new insights into cellular functions, disease mechanisms, and personalized medicine, heralding a more precise and comprehensive era in proteomics.

 

The applications of single-molecule proteomics are diverse and impactful. At the single-cell level, it allows for the detection of key protein molecules associated with diseases, the discovery of novel biomarkers, and the development of precise diagnostic tools for early disease detection. Single-molecule proteomics also facilitates the analysis of drug-target interactions, enabling the identification of effective and low-toxicity therapeutic targets. In studies of cellular heterogeneity, single-molecule proteomics reveals protein expression differences among individual cells, shedding light on the roles of heterogeneity in both physiological and pathological contexts. In immunology and tumor microenvironment research, single-molecule proteomics uncovers interaction mechanisms between immune and tumor cells, offering critical data to advance immunotherapy.

 

Principles of Single-Molecule Proteomics

The cornerstone of single-molecule proteomics lies in treating each protein molecule as a unique research subject. Using high-sensitivity detection techniques, single-molecule proteomics analyzes the sequence, structure, abundance, and PTM states of individual proteins. Key elements include:

 

1. Single-Molecule Isolation and Immobilization

Proteins must be isolated and immobilized on specific carriers to ensure individual analysis.

 

2. High-Sensitivity Detection

Detection requires exceptional sensitivity to capture subtle signals from single protein molecules. Common methods include single-molecule fluorescence microscopy, nanopore technology, and super-resolution imaging.

 

3. Sequence and Structural Analysis

By employing specialized chemical or physical methods, single-molecule proteomics determines the amino acid sequence and three-dimensional structure of proteins with high resolution.

 

4. Data Analysis and Integration

Single-molecule proteomics generates large, complex datasets, necessitating advanced bioinformatics algorithms for reliable and reproducible results.

 

Core Technologies in Single-Molecule Proteomics

1. Single-Molecule Fluorescence Microscopy

Fluorescent labeling of proteins enables high-resolution microscopes to detect and quantify signals from individual molecules, facilitating real-time observation of their dynamic behavior.

 

2. Nanopore Technology

Proteins are pulled into nanoscale pores using an electric field, with electrical current changes analyzed to decode sequences and PTMs. This label-free method directly reads amino acid sequences.

 

3. Single-Molecule Mass Spectrometry (SM-MS)

Combining high-resolution mass spectrometry with single-molecule analysis, SM-MS provides detailed insights into protein sequences, masses, and PTMs with precision and throughput.

 

4. Super-Resolution Microscopy

By surpassing the resolution limits of conventional optics, super-resolution microscopy captures fine structural details and spatial distributions of individual proteins.

 

Advantages of Single-Molecule Proteomics

1. Exceptional Sensitivity

Single-molecule proteomics detects proteins with minimal sample amounts, breaking traditional mass spectrometry's limitations.

 

2. Unveiling Protein Heterogeneity

Unlike traditional methods that average protein characteristics, single-molecule proteomics highlights unique molecular features, revealing true protein heterogeneity.

 

3. Real-Time Dynamic Tracking

Advanced imaging techniques allow for the observation of protein dynamics and spatial distributions in real time.

 

4. Comprehensive PTM Analysis

Single-molecule proteomics deciphers the PTM states of individual proteins, providing insights into regulatory mechanisms.

 

With its robust expertise and cutting-edge technology, MtoZ Biolabs offers tailored proteomics services for fundamental research, disease exploration, and drug target discovery.

 

MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.