Monoclonal Antibody Generation

  • Monoclonal antibody generation using immunization in mice
  • Hybridoma generation and selection
  • cDNA sequence  (DNA) coding for VH and VL and subsequently expressed on human IgG constant region
  • Humanization possible through a partner (Panorama Research Inc., Sunnyville, CA; Antitope Ltd, UK) and testing of various humanized versions of the antibody leads in functional potency assays

Cell Line Development

  • Cloning of DNA sequences in our CHOBC® vectors
  • Stable transfection of the proprietary CHOBC®cells
  • Selection of stable, high producing CHOBC® subclones
  • Upscaling of lead clones and securing 50 L lock process
  • Manufacturing of recombinant protein up to hundreds of grams
  • All relevant analytics and assays for protein characterization

Upstream Process Development

  • Stirred Tank Bioreactors ranging between 1 and 50 L scale
  • Modulation of the process to optimize titer and productivity (routine titers of 2 – 6 g/L)
  • Modulation of the process to target both the acidic and basic species dependent on the need
  • Mannosylation, galactosylation and fucosylation can either be increased or decreased

Downstream Process Development

  • Design of cost-effective and robust DSP processes capable of producing pharmaceutical-grade biotherapeutics
    • Optimization of cell clarification by depth filtration and sterile-grade filtration
    • Screening and optimization of various chromatography types
    • Set-up of normal flow and tangential flow filtration steps
    • Optimization of virus removal and inactivation steps, including in depth knowledge of viral validation studies
  • Antibody purification expertise: biosimilar and novel antibodies
  • Developing purification protocols for basically all protein-based therapeutics
  • Design of Experiment (DoE) solutions for process optimization and characterization
  • Extensive experience with disposable technology, including charged membrane filters
  • Scale up and technology transfer expertise

Analytical Development

  • Extensive experience in (e.g. mass spectrometry-based) analysis of complex proteins including Mabs
    • Peptide mapping, including full sequencing (UPLC-MS/MS)
    • Intact protein and semi-intact protein mass spectrometry (UPLC-MS/MS)
    • N-terminal and C-terminal protein sequencing (UPLC-MS/MS)
    • Deamidation, oxidation, pyro-glu (UPLC-MS/MS)
    • Determination of disulfide bonds (heterogeneity) (UPLC-MS/MS)
    • Released Glycans (UPLC-FLR-MS)
    • Determination of charge variants by Cation (CEX) and Anion (AEX)
    • Exchange Chromatography & by cIEF isoelectric focusing
    • Determination of aggregates and fragments by Size Exclusion
    • Chromatography (UPLC-SEC)
    • CE-SDS capillary electrophoresis
  • Determination of (CHO-based) Host Cell Proteins
  • High throughput MAb sample purification and sample analysis
  • Method qualification and validation
  • Technology transfer

(Bio)assay Development

  • Flow Cytometry
  • ELISA
  • Functional proprietary cell-based assays, including proprietary Fc all based domain binding assays
  • ADCC (using a proprietary effector cell line) and CDC
  • Cytokine release assays
  • Potency assays
  • Immunogenicity assays