Developing a safe and effective biotherapeutic depends on a thorough understanding of its critical quality attributes (CQAs), including protein integrity, glycosylation profiles, and post-translational modifications (PTMs).
At FyoniBio, we provide an in-depth characterization of your product’s protein and glycan profiles, helping you meet regulatory and quality control requirements across all stages of development and manufacturing. From conducting early development studies to regulatory-compliant validation, we design and adapt mass spectrometry assays to deliver dependable, reproducible results that support confident decision-making at every step.
Monoclonal Antibodies
Bispecifics
Antibody-Drug Conjugates
Fusion Proteins
Oligonucleotides
Glycoproteins
We apply bottom-up, middle-up, and top-down proteomics approaches for comprehensive characterization of your biotherapeutic.
We accurately determine molecular weight and assess the structural integrity of your protein, by identifying potential modifications, degradation, and truncations.
Using UPLC-MS/MS, we perform detailed analysis of proteolytic digests to precisely locate and quantify PTMs, confirm sequence integrity, and support batch-to-batch comparability studies for your protein.
We provide structural insights at the subunit level through the characterization of antibody fragments (Fab/Fc), heavy and light chains, and associated glycoforms—ideal for complex molecules and biosimilarity assessments.
We determine the drug-to-antibody ratio (DAR), assess conjugation sites, and evaluate the distribution and stability of your ADC product.
We verify correct disulfide bond formation and identify scrambled or incorrect linkages, ensuring proper folding and structural stability of your protein, while considering plausible modification of the affected (redox-sensitive) cysteines.
Solutions
Glycosylation plays a critical role in the stability, efficacy, and immunogenicity of biotherapeutics. Our high-resolution glycan profiling services help you characterize this essential quality attribute with precision and confidence—supporting development, comparability, and regulatory readiness.
We combine HILIC-UPLC-FLR with RapiFluor® or 2-AB labeling and our proprietary GlycoFiler® software to deliver automated, high-precision glycan profiling. Our MS-based glycan structure identification ensures accurate data you can rely on for regulatory submissions.
We apply 2-AB fluorescent labeling and HILIC-UPLC-MS/MS to analyze released O-glycans from proteins and peptides. This fast, efficient workflow minimizes N-glycan interference and accelerates your glycan analysis.
We provide site-specific insights into N- and O-glycosylation, including site occupancy and glycoform diversity. Our workflows support target-specific glycoform screening and help you map glycosylation heterogeneity across your molecule.
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Our analytical workflows are supported by advanced instrumentation and robust sample preparation methods, ensuring precision, reproducibility, and regulatory-grade data throughout your development pipeline.
Our advanced UPLC-MS platforms ensure exceptional accuracy, sensitivity, and reproducibility for your analytical requirements:
| Specification Category | System 1: Waters Acquity UPLC + PDA/FLR + Bruker Impact HD/II QTOF | System 2: Waters Acquity UPLC + FLR + Bruker Compact QTOF | System 3: Waters Acquity UPLC + FLR (Standalone) |
|---|---|---|---|
| Application Suitability | |||
| MS Detection Capability | MS, MS/MS (DDA, DIA) | MS, MS/MS (DDA, DIA) | – |
| Application Focus | Biomolecule characterization: intact protein mass, peptide mapping, disulfide linkages, oligonucleotides | N-/O-Glycan profiling, amino acid analysis, Biologic drug analysis, PK studies | Released glycan fingerprinting with fluorescent compound detection |
| Comments / Notes | High-resolution; ideal for detailed structural elucidation on intact level | High-performance MS; suited for routine analysis, quick and robust | No MS; ideal for fluorescence-based detection and chromatographic method development |
| Performance Specifications | |||
| Mass Range (Da) | 50 – 200,000 | 50 – 100,000 | – |
| Resolution | > 45,000 FWHM | > 20,000 FWHM | – |
| Mass Accuracy | < 0.5 ppm (internal calibration) | < 1 ppm (internal calibration) | – |
| Detector Type(s) | PDA, FLR, QTOF-MS | FLR, QTOF-MS | FLR |
Contact us today for a consultation or a personalized quote. Our team is here to support your goals with tailored analytical solutions.
We support compliance across all stages of development, from early discovery to post-market approval. Our validated methodologies and clear, well-structured documentation help you meet global regulatory requirements with confidence.
Every biologic is unique. That’s why we tailor our mass spectrometry strategies to the specific challenges of your molecule and your project’s needs.
Learn more about our glycan profiling capabilities.
Read about how our GlycoFiler® software can be used for automated glycan identification in mass spectrometric data analysis.
Posttranslational modifications are mostly connected to mass differences of the modified protein. These can be detected globally by intact mass measurement or site-specifically via peptide mapping.
Although most N- and O-glycopeptides can be detected through peptide mapping, the most sensitive and reliable detection of N-glycosylation is enzymatic release with N-glycosidase and fluorescence labeling. RapiFluor is a reliable and sensitive labeling reagent boosting both fluorescence and MS signal intensity. Fluorescent labeling with 2AB is available to analyze the O-glycosylation profile of proteins.
Most proteins are sensitive to oxidation of methionine and in rare cases also of tryptophan or histidine. Deamidation of asparagine and to a lower extent of glutamine is often seen as well as hydroxylation of proline and lysine. Further modifications are scrambling of disulfide bridges, glycation, sulfation, phosphorylation as well as terminal conversion or truncation.
Our experienced team is ready to provide the critical analytical support you need to make your drug development program successful.
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Illustration designer: Tilo Schneider
