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Metabolomics

Protein Full-length Sequencing

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Overview

Full-length sequence information is essential for studying the integrity and correct translation of biopharmaceuticals. In general, protein sequence analysis service provided by MtoZ Biolabs includes three steps: digestion of proteins into smaller peptide fragments; tandem LC-MS/MS analysis of peptides; MS/MS data analysis. Traditional protein sequencing only uses trypsin for digestion, which renders the final protein sequence coverage of around 60%. To ensure 100% protein sequence coverage, MtoZ Biolabs uses up to 6 kinds of protein enzymes for protein digestions, eliminating any missing peptides.

Service Workflow

Applications

• Determination of full-length sequence of Proteins/Peptides/Antibodies/Vaccines
Analysis of PTMs & chemical modifications on protein
• Verification of the correct translation of Recombinant Proteins

Sample Requirements

Format Gel bands, gel spots, and liquid samples are acceptable.
Quantity Gel bands/spots that are visible to naked eyes are adequate for protein identification. For liquid samples, a total of 5-10 ug proteins are required.
Purity Purity of liquid samples should be as high as possible. For liquid sample <10 ug, please avoid large amounts of detergent and salt ions, and note the buffer composition and the estimated amount of total proteins when submitting samples.
Note All reagents/solvent used must be of the highest purity to reduce contaminating substances. Samples should be handled with extreme caution and always in clean condition. Any source that may introduce contaminating proteins should be eliminated.

Case Study

In this project, we analyzed the full-length sequence of an antibody product sent from one of our customers. To ensure 100% sequence coverage, we selected 4 enzymes for protein digestion, based on the predicted antibody sequence. The sequence mapping results showed 100% sequence coverage.

Reports

• Experiment procedures
• Parameters of liquid chromatography and mass spectrometer
• MS raw data files
• Peptide identifications and intensity
• Protein identifications and intensity

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Address:
155 Federal Street, Suite 700, Boston, MA 02110, USA
Email:
info@MtoZ-Biolabs.com
Fax:
1-617-616-8054