Assay development

MFL Biologics has developed hundreds of assays for various types of analytical, bioassayor biosafetytesting purposes throughout drug discovery, development and manufacturing. Custom assays can be transferred from clients and further optimized and validated or we can work with clients to develop new assays by utilizing the technical expertise of our analytical, cell biology, virology, molecular biology, and microbiology laboratory staff.

Assays are developed and validated for bulk drug substance and final drug product in-process analysis and lot release(biosafety, purity, potency, identity, and concentration/titer). Using forced-degradation experimental designs during preformulation studies, stability-indicating methods are identified and optimized for use in product stability studies.

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Analytical Method Development

The Protein Analytical Sciences (PAS) department at MFL offers comprehensive analytical methodologies for assay development (Table 1) including extensive LC-MS capabilities. The PAS team supports a wide range of development activities including:

  • Cell line engineering
  • Upstream / cell culture development
  • Downstream purification development
  • Formulation development and analysis
  • Product biochemical and biophysical characterization
  • Biological characterization
  • Product comparability and similarity assessment
  • Writing of CMC sections for IND/BLA submissions
  • Analytical investigation/trouble shooting for GMP manufacturing related issues
  • Reference standard generation, characterization and management

Table 1. Examples of available methods
  • Protein concentration by UV280, etc.
Potency by bioassay and biological characterization
  • Cell-based or binding ELISA assays
  • Fc Functional assays
Physiochemical property
  • Aggregation (SEC)
  • Fragmentation (CE-SDS)
  • Charge variants (IEX, cIEF) and pI determination (cIEF)
  • Appearance, pH, osmolality
Process impurities/contaminants
  • HCP (ELISA), Protein A (ELISA), residue DNA (qPCR)
  • Endotoxin, bioburden, and sterility
Biochemical and biophysical characterization
  • Intact mass analysis (LC/MS)
  • Primary sequence analysis and PTMs (Peptide map HPLC/MS/MS)
  • Glycan profiling (HPLC or UPLC)

LC-MS Capabilities

  • LC-MS and LC-MS/MS technique to provide intact mass of proteins under reducing and non-reducing conditions, peptide mapping analysis
  • LC-MS and LC-MS/MS assays used in comparability studies, stability studies, and forced degradation studies to identify and characterize charge variants, impurities, and various modifications
  • Multiple MS systems available:
    • LTQ-Orbitrap Velos ETD MS
    • Thermo Q-Exactive MS
    • Waters Xevo G2 Q-TOF MS
    • Agilent 6530 Q-TOF MS
    • Agilent TOF