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Methods by Parameter

This section provides a convenient overview of our testing methods. Our methods are categorised depending on the respective testing parameter.

Identity

  • Amino Acid Analysis (AAA): Determination of amino acid composition of a protein. Used as confirmation of primary structure.
  • Carbohydrate analysis: N-linked oligosaccharide profiling by determination of the sialylated and desialylated carbohydrate structures
  • HPLC, Ion-exchange chromatography (IEC): Identity, content and impurity determination. Detection of oxidation and deamidation products.
  • HPLC, Reversed phase chromatography (RP-HPLC): Identity, content and impurity determination. Detection of oxidation and deamidation products.
  • HPLC, Size-exclusion chromatography (SEC): Separation of protein based on size. Determination of degree of aggregation
  • Isoelectric focussing (IEF): Determination of the isoelectric point (pl) of a protein and assessment of isoform distribution and impurity profiling.
  • MALDI TOF Mass spectrometry (MALDI-TOF MS): Confirmation of protein identity by peptide mass using theoretical AA sequence. Detection of post-translational modifications.
  • Optical rotation/ Polarimetry: Specific rotation of a pure material is an intrinsic property of that material at a given wavelength and temperature
  • Peptide mapping: Confirm identity/primary structure. Enzymatic digestion of protein to monitor lot-to-lot variability/post-translational modifications.
  • SDS-PAGE, reduced/ non reduced silver, CBB stain: Detection of┬ámono- and multi-meric forms of protein and impurities.
  • Sequencing, C-terminal: Determination of amino acid sequence from the C-terminus of protein. Used as a confirmation of primary structure
  • Sequencing, N-terminal: Determination of amino acid sequence from the N-terminus of protein. Used as a confirmation of primary structure
  • Spectrometry, Fluorescence
  • Spectrometry, IR
  • Spectrometry, UV-VIS: Determination of extinction coefficient and content of protein.
  • Thin-layer chromatography (TLC)
  • Western blot: Identification of proteins by their immunologic reaction with antibodies of known specificity

Physicochemical

  • Clarity/ Opalescence
  • Color, Achromicity
  • Loss on drying: Measuring moisture content in both high level and trace amounts in solids, liquids
  • Melting point
  • Osmolality/ Osmolarity: Osmolarity is the measure of solute concentration, defined as the number of osmoles (Osm) of solute per litre (L) of solution (osmol/L or Osm/L)
  • pH
  • Resistance to crushing
  • Visible particles

Purity

  • ELISA for HCP: Detection and quantitation of residual proteins including BSA, Protein A and host cell proteins. Proprietary HCP assays for CHO and E. coli; also useful in bioassays and protein characterization
  • ELISA, Leakage: To determine column leakage of e.g. Protein A, Protein G, Insulin and other ligands
  • HPLC, Ion-exchange chromatography (IEC)
  • HPLC, Reversed phase chromatography (RP-HPLC)
  • HPLC, Size-exclusion chromatography (SEC)
  • PCR for specific viruses: e.g. squirrel monkey virus
  • Phage determination: Determination of Phages
  • Q-PCR for residual DNA: Determination oft DNA
  • SDS-PAGE, reduced/ non-reduced silver, CBB stain
  • Spectrometry, Fluorescence
  • Thin-layer chromatography (TLC)
  • Western blot: Identification of proteins by their immunologic reaction with antibodies of known specificity

Impurity

  • ELISA for HCP
  • ELISA, Leakage
  • PCR for specific viruses
  • Q-PCR for residual DNA

Content

  • Carbohydrate analysis
  • ELISA: Detection and quantitation of Drugs substances
  • HPLC, Reversed phase chromatography (RP-HPLC)
  • Protein content (Bradford, BCA, Lowry, UV): ref. to parameter
  • SDS-PAGE, reduced/ non-reduced silver, CBB stain
  • Spectrometry, Fluorescence
  • Water content (Karl Fischer titrimetry)
  • Titrimetry (water-free): Determination of water (moisture) content

Functionality

  • Bioassay, Proliferation: Determination and quantitation of biological activity.
  • Bioassay, Differentiation
  • Bioassay, Cytotox
  • Bioassay, Apoptosis
  • Bioassay, Angiogenesis
  • Dissolution testing: Provide critical in vitro drug release information for both quality control purposes, i.e., to assess batch-to-batch consistency of solid oral dosage forms such as tablets, and drug development, i.e., to predict in vivo drug release profiles.

Safety

  • ELISA for (N)ADA: Detection and quantitation of antibodies against drug substance and drug product levels
  • PCR for specific viruses