Protein N-Terminal Sequencing Service

Understanding the structure of a protein's N-terminus is essential in biopharmaceuticals and protein engineering. N-terminal sequencing is the standard technique for verifying the boundaries of biologics and recombinant proteins, assessing protease-resistant regions, and identifying proteins in unknown species. This technique ensures the accuracy of protein synthesis and processing and characterizes the protein's functions, including detecting modifications like acetylation or phosphorylation, which affect stability and function.

 

Services at MtoZ Biolabs

MtoZ Biolabs offers high-precision N-terminal sequencing services utilizing high-resolution mass spectrometry and the gold-standard Edman degradation method. While Edman degradation confirms the identity and purity of synthesized or purified peptides and validates protein termini, mass spectrometry-based sequencing overcomes issues related to blocked or modified sites, providing comprehensive sequence coverage.

 



Figure1. Protein N-Terminal Sequencing Workflow Based on Mass Spectrometry and Edman

 

1. Mass Spectrometry-Based N-Terminal Sequencing

Proteins are enzymatically digested into smaller fragments. Each fragment's m/z and spectral data are analyzed to determine the N-terminal sequence, ideal for complex samples, including those difficult to isolate chemically.

 

Advantages

(1) Handles complex mixtures and extensive proteomic analyses.

(2) Identifies a broad range of chemical and biological modifications in proteins.

(3) Provides rapid and flexible analysis of minimal sample quantities.

 

2. Edman Degradation-Based N-Terminal Sequencing

This method involves the sequential chemical degradation of the protein's N-terminus, releasing one amino acid at a time, which is then analyzed chromatographically. This cyclic process precisely determines the entire N-terminal sequence, ideal for peptide identification and protein termini verification.

 

Advantages

(1) Offers high precision and repeatability, suitable for simple or purified protein samples.

(2) Independent of protein mass or complexity.

 

Sample Submission Requirements

1. Quantity: 5-10 micrograms of target protein.

2. Purity: Greater than 95% purity recommended.

3. Buffer Conditions: Avoid detergents or high salt concentrations.

4. Solvent: Use volatile solvents like water or acetonitrile, avoiding non-volatile solvents.

5. Storage: Store in low-binding microcentrifuge tubes, freeze at -20°C or -80°C, and transport with dry ice.

 

Applications

1. Protein Identification and Characterization: Confirm primary structure and N-terminal sequence.

2. Biopharmaceutical Quality Control: Ensure batch consistency and perform identity testing.

3. Post-Translational Modification Analysis: Identify modifications like acetylation, methylation, and cyclization.

4. Protein Engineering and Design: Confirm site-directed mutagenesis and conduct structural studies.

5. Biomarker Discovery: Diagnostic applications for early disease detection and monitoring.

6. Protein-Protein Interactions: Investigate interactions and drug development.

7. Structural Biology and Protein Folding: Explore protein folding dynamics and stability.

 

MtoZ Biolabs is dedicated to providing top-tier protein N-terminal sequencing services, tailored to meet specific research needs and commercial objectives. Our team offers comprehensive technical support and consultation to ensure precise utilization of N-terminal sequencing results in your projects. For further information or to consult about our services, please contact us anytime!