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Protein Characterization

Our team employs a customized approach to ensure successful generation of high-quality proteins with robust lot-to-lot reproducibility. Characterization is integrated throughout each project, rather than postponed until the end. This enables rapid orthogonal evaluation of critical protein-related key parameters. New assays are continuously implemented, and existing ones are adapted to meeting the specific needs of each application.

 

Orthogonal evaluation of related key parameters:
  • Purity - N-term/C-term homogeneity, chemical stability, post-translational modifications, residual protease identification
  • Size in solution - Oligomerization state, aggregation susceptibility, resistance to stress
  • Stability and folding - Low and high order structure, thermodynamic evaluation, forced degradation study
  • Sequence integrity - N-term/C-term homogeneity, chemical stability, post-translational modifications
  • Biomolecular interaction - Rate of association/disassociation
 
 

Characterization and analytical methods

 

Protein concentration

  • Colorimetric (Bradford/Lowry/BCA/RCDC)
  • Spectrophotometric: absorbance at 280nm

Aggregation and sizing

  • SEC-HPLC / SEC-MALS
  • Analytical ultracentrifugation (SV-AUC)
  • Dynamic light scattering (DLS)
  • SDS-PAGE
  • Glutaraldehyde crosslinking

 

Reactivity

  • ELISA

Purity

  • Analytical SEC-HPLC
  • Capillary electrophoresis
  • SDS-PAGE/ densitometry
 

Identity

  • Western blot
  • Peptide mapping (LC-MS/MS)
  • Dynamic light scattering

 

 

 

Secondary / tertiary structure

  • Circular dichroism (far/near UV CD)
  • UV/Vis absorbance spectroscopy 
  • Intrinsic fluorescence
  • Differential scanning fluorimetry and calorimetry
  • Transmission electron microscopy
  • N-glycan analysis

Intact molecular weight

  • SEC-MALS (multi-angle light scattering)
  • LC-MS

 

Process related impurities

  • Residual proteins; residual DNA (qPCR)
  • Enotoxins (LAL)
  • Sterility

Protein activity

  • Protein binding affinity (Bio-layer interferometry)
  • Enzymatic/colorimetric assays
  • Capillary electrophoresis
  • SDS-PAGE/ densitometry
 

Thermal stability

  • Differential scanning fluorimetry (DSF) and calorimetry (DSC)

 

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