Putting Science to Work
Protein Sciences

Protein Sciences

Protein Sciences

Syngene's comprehensive gene-to-protein services start with cloning the gene and extend up to the supply of ready-to-use proteins. Proteins can be supplied as per the specific needs of the client - ranging from mg to gram scale and varying from research grade to highly pure crystallography grade.

A typical gene-to-protein program includes codon optimization and synthesis, screening of expression in different host systems (bacteria, yeast, insect cells, mammalian cells), expression optimization, expression scale up, purification and quality analysis.

Protein Expression

Expression in Bacteria

  • Bacterial hosts such as E.coli, Bacillus
  • Expression screening in E.coli to identify suitable host strain and optimal conditions for expression
  • Scale of expression: 2 - 10 L lab scale and 20 - 225L pilot scale

Expression in Yeast

  • Hosts such as Saccharomyces sp. and Pichia pastoris
  • Expression screening to identify suitable host strain and optimal conditions for expression
  • Scale of expression: 2 - 10 L lab scale and 20 - 225 L pilot scale

Expression in Insect cells

  • Insect Cell Lines used: Sf9, Sf21, High Five
  • Generation of high titer recombinant baculovirus
  • MOI studies
  • Scale of expression: 2 to 50L in Wave Bioreactor

Expression in mammalian cells

  • Mammalian Cell lines: CHO-K1, HEK 293, CHO-S, CHO-DG44 etc.
  • Transient transfection for expression and purification of proteins
  • Generation of stable cell lines expressing recombinant proteins
  • Scale of expression: 2 - 10 L lab scale and 30 - 300 L pilot scale
Protein Purification
    • Use of state-of-the-art chromatography systems (Akta)
    • Purification of mg to gram quantities of proteins for various applications (research, assay, animal studies, crystallography)
    • Development and optimization of purification strategy using multiple steps of chromatographic methods:
      • Affinity
      • Hydrophobic
      • Ion exchange
      • Size exclusion

  • Purification of recombinant proteins containing designed purification tags : His6, GST, MBP, SUMO, FLAG, etc.
  • Option of tag removal by proteolytic digestion (Thrombin, TEV, PreScission, SUMO protease, Factor Xa, etc.)
  • Purification from inclusion bodies:
    • Development of refolding methods by exhaustive screening of refolding conditions
    • Refolding scale up and purification of properly folded protein
Protein Characterization, Analysis and Quality Control

A myriad of techniques are employed to analyse each batch of protein produced that ensures strict quality control and minimal batch to batch variation:

  • Electrophoresis (SDS-PAGE, native-PAGE)
  • Capillary Gel Electrophoresis (CGE)
  • DOT/Western blot
  • Imaging Capillary Electrophoresis (iCE)
  • Mass Spectrometry (ESI-MS/MS, MALDI-TOF/TOF)
  • Peptide Mass Finder Printing
  • Reverse Phase-HPLC
  • Size Exclusion HPLC
  • Glycoform Analysis
  • Endotoxin Level Detection
  • Biological Activity Assays
Protein Crystallography

Syngene's protein crystallography services aim at providing one-stop-solution for gene-to-structure programs. In collaboration with the strong Bioassay and Medicinal Chemistry groups of Syngene, the crystallography facility is designed to support any structure-based drug design program. The crystallography services offered are:

  • Crystallization screening and optimization
    • Commercially available screens
    • Customized screens
    • Sitting drop, hanging drop, microbatch (under oil) and seeding (Mosquito crystallization robot)
    • Microscopic examination of drops, Staining, Mass spectroscopy of harvested crystals (MALDI)
    • Electrophoresis, preliminary X-ray diffraction
    • Custom screening
    • Crystallization of protein-inhibitor/ligand complexes (co-crystallization)
    • Soaking of crystals in inhibitor/ligand solution

  • Cryoprotectant screening by preliminary X-ray diffraction
  • Complete X-ray data collection using rotating anode source
  • Synchrotron data collection (mail-in format)
  • Molecular replacement and structure refinement
  • Structure analysis and data presentation

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