Confarma

Protein characterization

Solvias provides comprehensive cGMP services at every stage of drug development. We can fully characterize any protein biopharmaceuticals for the Chemistry and Manufacturing Controls (CMC) section of Investigational New Drug applications (INDs), New Drug Applications (NDAs) and Biologic License Applications (BLAs).

According to the ICH Q6B guideline, the analytical characterization of a protein should include primary, secondary, tertiary, and quaternary structural assessment, biological activity, and analysis of product and process impurities. Therefore, Solvias typically performs:

Structural Characterisation

  • Amino acid composition
  • Primary sequence analysis including N- and C-terminal truncation (peptide mapping by proteolytic digestion and LC-MS/MS, Edman degradation)
  • Peptide mapping
  • Amino acid misincorporation analysis
  • Free sulfhydryl group and disulphide bridge analysis (mis-matched bridges)
  • Carbohydrate structure determination: carbohydrate content (neutral, amino sugars, sialic acid), glycosylation site determination, antennary profile, glycan composition analysis
  • Post translational modification analysis by HRMS (Orbitrab): Deamidation, Oxidation,
  • Glycation, Glycosylation, Phosphorylation, Scrambled disulfide bridges, Truncation, N- and C-terminal modifications

Physico-chemical Analysis

  • Molecular weight by Mass Spectrometry (MALDI-MS and Orbitrap LC-MS)
  • Isoform and impurity studies using Chromatographic (IEX, SEC, RP, HIC, HILIC, NP, HPAEC, IC, Affinity Chromatography ) and Electrophoretic methods (PAGE, SDS-PAGE, IEF, CGE, CIEF, CZE, iCE280, Bioanalyzer)
  • Extinction Coefficient determination
  • Liquid chromatographic profiles identification, heterogeneity, purity (HPLC, UPLC, IEX, SEC, RP, HIC, HILIC)
  • Quantity by UV Absorbance at 280 nm, Bradford Assay, Quantitative amino acid analysis, BCA

Spectroscopic Profiles

  • UV-VIS
  • Circular Dichroism (CD) (for secondary and higher order structure)
  • Nuclear Magnetic Resonance Spectroscopy (NMR)
  • Infra Red (FTIR) Spectroscopy
  • Fluorescence Spectroscopy

Aggregation Studies

  • Analytical ultracentrifugation, SEC, SEC-MALLS, DLS, Zeta Potential

Solvias analytical methods for biologics characterization includes:

  • HPLC methods: IEX, SEC, RP, HIC, HILIC, NP, HPAEC, IC, Affinity Chromatography
  • HPLC detection: UV, FL, RI, ELSD, CAD, MALLS, MS
  • Electrophoresis: Slab gel (SDS-PAGE), CGE, CIEF, CZE, iCE280, Bioanalyzer
  • Spectroscopic methods: CD, UV, FL, IR, NMR, DLS, MALLS, DSC
  • Mass spectrometric methods: MALDI TOF-TOF, LC-MS(n) (Orbitrap, Triple Quad, Ion Trap)
  • Others: FPLC, AUC, AF4, Immunological methods, ELISA, Western blotting, Residual DNA (threshold analysis, qPCR), Rodent parvoviruses, Bioburden, Endotoxin, Sterility, Potency assays