Yaohai Bio-Pharma offers N-terminal sequence testing and data analysis services based on Liquid Chromatography-Mass Spectrometry (LC-MS) or Edman degradation method, which allows you to determine the amino acids at the respective termini of protein of interest. N-terminal modification can also be analyzed by LC-MS.
We have been involved in the Protein Structural Characterization of various large molecules, including Recombinant Subunit Vaccines, Nanobodies/VHHs/ Single-domain Antibodies (sdAbs), Antibody Fragments, Hormones/Peptides, Cytokines, Growth Factors (GF), Enzymes, Collagens, etc.
Regulatory Requirements for N Terminal Sequencing
According to the ICH Q6B guidelines, it is necessary to confirm the sequence of your protein and examine the termini for any variation that may exist.
N Terminal Sequencing Methods
| Analysis |
Method |
| N-terminal sequencing |
Edman degradation, Liquid Chromatography-Mass Spectrometry (LC-MS) |
Procedures
LC-MS
1. Protein N-terminal fluorescent labeling
2. Multiple digestions
3. LC-MS/ Mass Spectrometry (MS) analysis
Edman degradation
1. Sodium Dodecyl Sulfate (SDS) gel electrophoresis of your target protein
2. Transfer gel to Polyvinylidene Fluoride (PVDF) membranes and staining
3. Identify and cut target protein band
4. Edman sequencing of N-terminus
5. Data analysis
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