CMC Support Services

Consistency and safety of manufactured pharmaceutical or biologic chemical entities requires detailed physicochemical characterization. Improving and de-risking the overall drug development path from Phase I clinical trials to approval requires advanced analytical assessment of critical quality attributes.

CMC (Chemistry, Manufacturing, and Controls) evaluation of biological compounds, including antibodies, occurs at various stages of development and involves monitoring a range of critical quality attributes through peptide mapping, the determination of disulfide linkages, localizing post-translational modifications, and glycan profiling.* Modern biopharmaceutical drugs, such as recombinant proteins, monoclonal antibodies (mAb), bispecific antibodies, vaccines, antibody drug conjugates (ADC’s), oligonucleotides and gene therapy products (e.g. AAV’s) are very complex molecules. Characterizing and monitoring these compounds for CMC and other quality control environments can present a significant challenge. Our CuriLytics™ platform can help you solve your most complex analytical challenges by designing flexible and customized programs that encompass complete characterization according to ICH Guideline Q6B (Appendix 6.1).

Structural Characterization

  • Amino acid sequence
  • N- and C- terminal sequence
  • Peptide Mapping
  • Free Sulfhydryl groups and disulfide bridges (disulfide bonds)
  • Monosaccharides and carbohydrate structure
  • Spectroscopic profiles

Purity/Product related impurities

  • Degradation
  • Aggregation
  • Oxidation
  • Deamidation

Physicochemical Properties

  • Molecular weight and size
  • Isoforms and Isoelectric point (charge heterogeneity)
  • Extinction Coefficient
  • Glycan analysis
  • Spectroscopic profiles

Process related impurities

  • Residual Solvents
  • Residual DNA
  • Residual Host cell protein
  • Detergents
  • Endotoxin

Amino acid sequence

Peptide mapping is not only used to confirm the amino acid sequence of a biological chemical entity (protein), but also characterizes post translational modifications including phosphorylation, deamidation, oxidation, fragmentation, and glycosylation, sulfation, phosphorylation, or formylation. Proteins are reduced, alkylated, and enzymatically digested. We optimized the range of digestion enzymes used to maximize coverage of your protein using high resolution mass spectrometry (RP LC/MS/MS). This also allows confident confirmation of protein amino acid sequence, details any sequence variants, confirms N- and C-termini sequence, and details post-translational modifications.

Intact protein molecular weight

Obtain accurate molecular weight of biologics using state-of-the-art high resolution mass spectrometry (LC/MS). Characterize the post translational modifications of a protein including glycosylation, oxidation, phosphorylation, deamidation, etc.. In combination with peptide mapping (used in Amino Acid Sequence analysis) this allows identification and localization of a range of post translational modifications on your protein at both the protein and peptide level. Additionally, SEC-HPLC and SEC-HRMS can accurately detail the mass and size heterogeneity of your protein.

Molecular weight of heavy and light chain of antibody

High resolution mass spectrometry provides accurate molecular weight of reduced antibody biologics. The independent measurement of the molecular weight of both light and heavy chains provides additional confirmation of the amino acid sequence of an antibody and provide a more detailed assessment of post translational modifications on each chain.

Disulfide linkage

Disulfide bonds are an important mechanism used to stabilize protein structures. Breaking or scrambling of disulfide bonds is strongly associated with loss of protein function and activity. We utilize high resolution mass spectrometry and an optimized range of digestion enzymes to map disulfide bonds of proteins and characterize any disulfide scrambling.

Glycosylation and glycan profiling

Glycosylation plays a critical role in determining protein structure, function, and stability. Identifying the glycans linked to a protein is therefore important in assessing analytical aspects of a glycoprotein. The glycosylation profile for a protein can be obtained with glycans attached to the protein and/or when glycans are enzymatically released from the protein. Analysis of the glycans released from a glycoprotein can serve to confirm glycoprofiles produced from intact protein LC/MS analysis

Glycan profiling of intact proteins and antibodies

    • The procedure can identify glycans on any glycoprotein, including antibodies.
    • This procedure uses high resolution mass spectrometry (MS) detection

Glycan profiling through released glycan analysis

    • This protocol identifies glycans that are enzymatically removed (“released”) from a glycoprotein
    • The procedure identifies n-linked glycans but not o-linked glycans
    • This procedure uses both high resolution mass spectrometry (MS) and fluorescence detection

Charge heterogeneity

We utilize high resolution mass spectrometry, CE, IEF to analyze proteins to accurately determine charge variants in biologics.

*GUIDANCE FOR INDUSTRY FOR THE SUBMISSION OF CHEMISTRY, MANUFACTURING, AND CONTROLS INFORMATION FOR A THERAPEUTIC RECOMBINANT DNA-DERIVED PRODUCT OR A MONOCLONAL ANTIBODY PRODUCT FOR IN VIVO USE, section A.2., Center for Biologics Evaluation and Research (CBER), Center for Drug Evaluation and Research (CDER), August 1996