Introduction
Asparagine (Asn) deamidation and aspartic (Asp) isomerization are two of the most common degradation reactions that
occur during long-term storage of mAb products, and they can occur at different stages of the mAb production
process. Product heterogeneity resulting from uncontrolled degradation due to deamidation and isomerization can
complicate manufacturing consistency, hampering long-term mAb functionality. Therefore, drug candidates prone to
degradation should be identified early in the drug development process to adjust manufacturing and formulation
processes accordingly or redesign problematic drug candidates to eliminate such hotspots.
Fig.
1. Potential chemical degradation sites (deamidation and isomerization) in an antibody (PDB: 1IGT). (Kuroda D,
et al., 2020)
Our Services
Accurate assessment of chemical modifications such as asparagine deamidation and aspartate isomerization is an
important part of the comprehensive characterization of therapeutic monoclonal antibodies (mAbs). When located in
complementarity-determining regions (CDRs), these modifications lead to loss-of-function, affecting product efficacy
and safety, and leading to the designation of the modification as a critical quality attribute.
As a quality partner of the biopharmaceutical industry. Creative Proteomics has successfully
completed many challenging projects in antibody analysis. You have reason to believe that we are the ideal partner
for your project.
Creative Proteomics offers a wide range of services for the characterization of therapeutic
proteins. Whether you're looking for a custom or off-the-shelf solution, Creative Proteomics
has over years of expertise in delivering unique and powerful assays to help accelerate your treatment plan.
Here, our experienced scientists have developed an advanced platform for antibody deamidation and isomerization
analysis. Our antibody deamidation and isomerization analysis services are based on peptide mapping analysis. To
accurately characterize antibody deamidation and isomerization, Creative Proteomics' expert
team uses the following protocol.
- The monoclonal antibody is enzymatically digested with an endoprotease such as trypsin, which selectively
cleaves the peptide bond at the C-terminus to lysine and arginine residues (but not before proline).
- The resulting proteolytic peptides are separated and identified by liquid chromatography-mass spectrometry
(LC-MS) using accurate mass measurement.
It is noted that to enhance the characterization of sequence responsibility, liquid chromatography coupled with
online tandem mass spectrometry (LC-MS/MS) is required to provide site mapping of chemical modifications, as well as
direct, confirmatory sequencing for peptide identification. Proteolytic digestion is usually performed overnight at
the optimal pH for the enzyme to ensure complete digestion of the protein.
In addition to clear, easy-to-understand reports, you can expect:
- Expert advice direct from our experienced technicians.
- Quick turnaround, results typically within a week.
- Validation studies with application reports are available upon request.
Creative Proteomics is a reliable biopharmaceutical partner. Our professional team can provide
professional, systematic antibody deamidation and isomerization analysis service for our clients worldwile. Our
services guarantee accurate and reliable results, at quick turnaround time! If you would like more information about
specific aspects of our services, feel free to contact us.
References
- Kuroda D, Tsumoto K. (2020) Engineering Stability, Viscosity, and Immunogenicity of Antibodies by Computational
Design. J Pharm Sci. 109(5):1631-1651.
