C-Terminal Sequencing Service
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The C-terminus is an important structural and functional site of proteins, and some specific post-translational modifications such as C-terminus shearing affect the protein's structure and thus its biological function by affecting its structure. Therefore, C-terminus sequencing is critical for the stability of antibody drugs, their folding, and their interaction with receptors. Creative Proteomics provides professional C-terminal sequencing services to efficiently determine the C-terminal amino acid sequence of antibody proteins, providing valuable information for antibody drug manufacturing, quality control and regulatory requirements.
Importance of C-Terminal Sequencing in The Primary Structural Characterization of Antibody Drugs
C-terminal sequencing is a critical step for the consistency and quality of antibody drugs. The role of this technology in the characterization of antibody drugs includes the following:
- C-terminal sequencing effectively identifies any changes and modifications in the C-terminal region, and provide critical information about the integrity and homogeneity of antibody drugs.
- The composition of certain amino acids in the C-terminal region affects the antibody's efficacy, binding affinity, and signaling pathway. C-terminal sequencing can facilitate the understanding of the potential function and activity of antibody drugs and accelerate the further development of drug candidates.
- C-sequencing plays a crucial role in regulatory submissions and intellectual property protection. Having a comprehensive understanding of the C-terminal region allows for detailed characterization and differentiation of antibody drugs.
Antibody Drug C-Terminal Sequencing Platform
To evaluate the C-terminus, Creative Proteomics scientists use carboxypeptidase digestion or mass spectrometry analysis strategies to provide accurate and important information about the C-terminal sequence of antibody drugs.
Carboxypeptidase digestion
Carboxypeptidase digestion is a commonly used method for the C-terminal sequencing of antibody drugs. This technique involves the selective cleavage of amino acids from the C-terminus of the antibody protein using the enzyme carboxypeptidase. The process involves denaturing the antibody using heat or chemical reagents to expose the C-terminus. Carboxypeptidase is then added to the denatured antibody solution for digestion. The enzyme effectively removes the terminal amino acid residue from the C-terminus, leaving a new C-terminal amino acid. After digestion is complete, the released amino acids are identified and quantified by our advanced high performance liquid chromatography (HPLC) or mass spectrometry (MS) techniques.
Mass Spectrometry C-Terminal Sequencing
Mass spectrometry digests antibody proteins into short peptide fragments, which are then detected by LC-MS/MS analysis. The integrity of the C-terminus or other mutations of the antibody protein can be effectively assessed by comparing the data obtained from the peptide map with the expected antibody sequence. This method avoids the interference of C-terminal closure or modification sites and can detect post-translational modifications (e.g. phosphorylation, glycosylation, etc.) of antibody proteins, which can be used to assess the function and stability of antibody proteins.
C-terminal sequencing technology process at Creative Proteomics
The C-terminal sequencing process typically includes the following steps:
Antibody drug samples are denatured and digested enzymatically to release C-terminal amino acid residues.
- Separation and purification
Depending on the hydrophobicity of the peptide fragments, liquid chromatography techniques are used to separate the generated peptide fragments.
- Analysis and identification
The isolated peptide fragments were then analyzed and detected by LC-MS/MS.
The obtained mass spectrometry data is analyzed and interpreted to determine the amino acid sequence of the C-terminal end of the antibody drug. Database searches and specialized software tools are utilized to match the experimental data with known amino acid sequences.
Creative Proteomics provides timely C-terminal sequencing of your antibody drugs and comprehensive data analysis reports through our advanced technology to help you confirm whether your antibody protein drugs are fully expressed or otherwise mutated. Contact us to learn more about C-terminal sequencing service for antibody drugs. We will be happy to assist you.
