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Protein Sciences and Structural Biology

Protein Sciences and Structural Biology

Designed to meet scale and fit-for-purpose quality criteria of tool and therapeutic molecules.

Augmented by protein X-ray crystallography capabilities to drive integrated structure based discovery programs.

Our team has experience in handling difficult and complex target molecules. We offer wide range of services and solutions with the flexibility of multiple entry points. We are proficient with most modern expression platforms and our facilities are equipped to handle scales ranging from mgs to grams.

Protein crystallography services provide a one-stop solution for determining 3D structure of: Proteins and complexes : Protein-protein complexes Protein-small molecule complexes Antibody-antigen complexes at high resolution

Protein crystallography projects are typically carried out in gene-to-structure format encompassing gene cloning, protein expression, purification, crystallization, screening, synchrotron x-ray diffraction, structure solution and refinement.

Our structural biology expertise also includes protein engineering to enhance crystallizability of difficult proteins. We seamlessly integrate structural biology with drug discovery programs for end-to-end SBDD offerings.

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Expression Systems

Mammalian: Mammalian systems used for expression are CHO, ExpiCHO, HEK, ExpiHEK, etc. Protein production in transient expression mode or using stable clones or pools.

Insect cell: Baculovirus mediated protein expression and production in Sf9, Sf21, Hi5 and super Sf9 insect cell lines

Yeast: Stable protein expression in Pichia, recombinant molecule is typically secreted in media by genome integration.

Bacterial: Soluble, periplasmic and inclusion bodies in E. coli. Multi-protein expression by co-transfection or duet plasmid technology. Refolding condition screening and method optimization. Construct design, scouting and screening studies for challenging proteins. Protein expression scale-up using spinner flask, shaker flask, HYPER flask and WAVE bioreactor platforms to handle volumes up to 50 L scale with low operating costs.

Protein Purification

Combination of high-end instruments and a wealth of scientific expertise we provide the best in class customized purification platforms for research and drug discovery needs. We are proficient in purifying tagged as well as un-tagged and all classes of target proteins from mammalian, yeast and microbial expression systems.

  • Purification process optimization
  • Protein purification in batch binding mode as well as automated FPLC systems
  • Chromatographic purification methods include IMAC, IEC, mixed-mode, HIC, SEC and affinity for micrograms to gram-scale quantities

Antibody Purification

  • Purification of monoclonal antibodies of different origins (human, non-human primates, mouse, rat, rabbit, pig, hamster, gerbil, etc.)
  • Purification of antibody fragments of various formats (Fab, Fab2, F(ab)’2, VHH, scfv and generation from full length mAbs)
  • Bi-specific and KIH antibodies
  • Diabodies
  • Single domain antibody
  • Protein purification from hybridoma cultures

Microbial Purification Capabilities

  • Soluble protein
  • Periplasmic protein
  • Intracellular protein
  • Refolding from inclusion bodies
  • Purification of Major Histocompatibility (MHC) complexes
  • Membrane protein production – GPCRs, nanodisc technology, etc.
  • Antibody drug conjugate screening, development and purification
  • High throughput protein screening and purification
  • Protein cleavage and purification

Protein Analytics and Characterization

We are committed to deliver high-quality products by performing a number of routine protein analyses throughout all project stages ensuring the identity, purity, structural and conformational integrity, stability and function of the protein.

  • Quantitative analysis: Protein estimation using Absorbance at 280nm, Bradford and BCA assays
  • Qualitative analysis: SDS-PAGE (Reducing/Non-reducing) and DOT/Western blot analysis
  • Purity: SE-HPLC, IEX-HPLC and RP-HPLC
  • Protein stability analysis (Real-time and Accelerated Stability Testing)
  • LC-MS analysis
  • Capillary Electrophoresis (CE)
  • Peptide Mass Fingerprinting (PMF)
  • Biological activity and binding studies using enzymatic assays, ELISA, BIAcore etc.
  • Endotoxin analysis
  • Host Cell Protein (HCP) and Host Cell DNA (HCD) analysis

Protein Stability Analysis

  • Real-time and accelerated stability testing
  • Long-term stability studies

Protein Conjugation and Labeling

  • Protein labeling/conjugation (HRP, biotin, Ruthenium, fluorescent labels etc.)
  • Conjugation of resin with specific protein/peptide

Crystallization Screening

  • Multi-dimensional optimization of crystallization conditions using commercial and customized screens for a target protein or complex
  • Sitting drop, hanging drop, micro-batch (under oil) and seeding
  • Micro-seeding and macro-seeding

Crystal Hit Identification

  • Microscopic examination of drops, staining and mass spectromety of harvested crystals
  • Preliminary X-ray diffraction

Protein-protein and Protein-drug Complexes

  • Crystallization of protein-inhibitor/ligand complexes (co-crystallization)
  • Soaking of crystals in inhibitor/ligand solution

Data Collection and Refinement

  • Cryoprotectant screening
  • Complete X-ray data collection using rotating anode source
  • Synchrotron data collection at Australian synchrotron
  • Molecular replacement and structure refinement
  • Structure analysis and data presentation

Why Aurigene Pharmaceutical Services?

Multidisciplinary team of scientists

Scalability with high purity and exceptional quality

Processing complex molecules

Protein engineering for enhanced crystallizability

Synchrotron data collection

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