N/C Terminal Sequencing

Overview

N- and C-terminal sequences are important structures and functional parts of protein and polypeptide, and they may play decisive roles in the biological function of protein. The main methods for protein C-terminal sequencing include carboxypeptidase, chemical and tandem mass spectrometry, and the common methods for N-terminal sequencing include Edman degradation sequencing and mass spectrometry. Each of these methods has the respective advantage and shortcoming. Therefore, the combination of a variety of different sequencing methods can adapt to the requirements of multiple protein sequencing. For example, Edman degradation may not well solve the problem of blockage and protein modification on the N-terminal end. This drawback can be overcomed by mass spectrometry analysis. MtoZ Biolabs uses both Edman degradation system and MALDI-TOF for N/C sequencing of biopharmaceutical products. This combination of two methods ensures accurate analysis of N-terminal blockage and modified protein N/C terminal.

MtoZ Biolabs uses both Edman degradation system and MALDI-TOF for N/C sequencing of biopharmaceutical products. This combination of two methods ensures accurate analysis of N-terminal blockage and modified protein N/C terminal.

Biopharmaceutical N/C-terminal Sequencing

Sample Requirements

Sample types: lyophilized & solution samples
Amount of total protein: >300 ug
Purity: >90%
Content of salt: Volatile salt<200 mM; Non-volatile salt<5 mM

Case Study

In this case, the N- and C-terminal sequence of one of our clients’ protein sample has been analyzed using a combination of Edman degradation and MALDI-TOF technologies. The sequencing data highlighted in red color accurately shows the N- and C-terminal sequence of the protein.

Reports

• Experiment procedures
• Parameters of liquid chromatography and mass spectrometer
• Edman sequencing and MS raw data files
• Biopharmaceutical N/C-terminal sequence results
• Bioinformatics analysis

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Identification of Biopharm

Molecular Weight Analysis Peptide Mapping Amino acids composition

Variation Analysis

Glycan Profiling Glycosylation site analysis Disulfide bond analysis Antibody C-terminal variation analysis

Purity Analysis

Purity analysis SDS-PAGE Purity analysis SEC and RPLC Host Cell Protein (HCP) Analysis Antibody-Drug Conjugate (ADC) Analysis