This method determines the exact molecular weight of intact glycoproteins, especially antibodies, without requiring enzymatic digestion or fragmentation.
Depending on the project’s requirements, we perform this analysis using LC–MS or MALDI–TOF. When needed, proteins can be treated with specific glycosidases before intact mass measurement to confirm the nature and extent of glycosylation.
Intact mass analysis of purified protein by MALDI-TOF.
Purposes and applications for Intact Mass Analysis
Intact Mass Analysis of a purified glycoprotein serves several purposes and applications:
Oncology
Intact mass analysis is particularly valuable for characterizing therapeutic antibodies and cancer biomarkers.
Monoclonal antibodies’ (mAbs) efficacy and stability depend heavily on their precise molecular structure. Intact mass analysis allows:
- The identification of protein variants or degradation products
- Confirmation of the complete molecular weight
- Detection of PTMs
Infectious Diseases
- Analyze bacterial and viral proteins, including vaccine antigens and virulence factors
- Monitor structural variants of viral envelope proteins, such as those from influenza or SARS-CoV-2, which are prone to glycosylation changes that affect immune recognition
- Confirm the integrity and uniformity of glycoconjugate vaccines, which are widely used in infectious disease prevention
Immune and Inflammatory Diseases
- For therapeutic antibodies or those produced in response to infection or vaccination, determining the exact mass allows for the detection of subtle differences caused by isotype switching, glycosylation changes, or fragmentation
- Mass profiling of cytokines or immune receptors helps researchers study immune dysregulation in diseases such as autoimmune disorders or chronic inflammation
Mircrobiome
Enabling direct profiling of microbial proteins or excreted glycoproteins that interact with the host. It may be useful in the quality control of engineered probiotics or microbial therapeutics, ensuring that expressed proteins are intact and properly folded
Biomarkers & Diagnostics
By comparing the intact mass profiles of proteins from healthy individuals and patients, researchers can detect disease-specific isoforms in disease-related changes in large, glycosylated plasma proteins. Unlike peptide-based approaches, here we can monitor full-length protein forms, including PTMs, truncations, and glycoforms, which are critical to biomarker function and specificity
Other Fields
- Plant science and agriculture: It helps confirm the identity and integrity of defense-related proteins and enzymes
- Cosmetics industry: the technique ensures the purity and structural consistency of peptides and proteins used in skincare and haircare products
- Nutraceutical: to assess proteins, peptides, or sugars from natural sources, supporting quality control and bioactivity claims. It is also essential in the analysis of HMOs, allowing accurate profiling of these complex sugars
