Recombinant Proteins

Protein Target Development

• Custom gene synthesis and codon optimization
• PCR amplification of encoding sequence from genomic or plasmid DNA
• Proteins can be designed with "HIS-TAGS", "GST-TAGS", or client selected

Expression Systems

• Selection of appropriate expression vector and insertion of target sequence
• Transfection of host cell lines including bacterial, yeast, mammalian, and insect cells
• Pichia expression
• Small scale fermentation to assess expression levels and characterize protein
• Cell lines include
  • Mammalian
  • Bacterial
  • Yeast
  • Insect

Purification of Recombinant Proteins

• Development of purification protocols
• Cell lysis and precipitation
• Chromatography and polishing chromatography
• Purification to > 95%

Characterization of Recombinant Proteins

• Protein sequencing and mass spectrometry
• Mass spectral analysis and mapping my MALDI-TOF/MS, ES/MS, PDS/CID/CAF
• ELISA
• SDS-Page and Western Blots
• Amino acid analysis
• Stability Testing

Biophysical Characterization

• Isothermal titration calorimetry (ITC). Determination the binding constant and stoichiometry of binding between molecules, as well as other thermodynamic parameters such as the molar heat, free energy, and entropy of binding.
• Thermal stability characterization by differential scanning calorimetry (DSC)
• Structural analysis by Circular Dichroism, Florescence Spectroscopy, Protein Florescence, Light Scattering Spectroscopy (SEC)

Scale Up Fermentation