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Reagent
Generation

Aragen is a valued partner for gene to protein studies, offering specialized support from custom study design, drafting and implementation of workflows, to delivery of reagents. We have a solid track record of purifying 200+ functionally active recombinant proteins and generating 100+ cell lines as stable or transient expression systems for drug candidate screenings. The integration of molecular biology, cell culture, purification and product analysis allows us to deliver complete packages of purified products, with associated functional and analytical data.

We also support structural and functional assays, target validation and throughput screens with extensive expertise in the production of recombinant proteins, monoclonal and recombinant antibodies, vaccine antigens and disease biomarkers as customized reagents. A dedicated project management team monitors progress and communicates in real time, offering clients flexibility throughout the process.

Molecular Biology

Our molecular biology team offers a spectrum of services including gene knockout/knockdown with advanced TALEN-Fok1/CRISPR/Cas9 technologies, reporter gene construct generation and dual promotor (bi-cistronic) harboring expression construct generations. We facilitate fully customized expression constructs to provide custom solutions that meet our customers’ research goals.

  • DNA
    • codon Optimization
    • DNS synthesis from protein sequences
  • Gateway cloning
    EXPRESSION
    CONSTRUCT
    GENERATION
    • Expression construction generation
    • DNA sequencing and analysis
    TOPO/RE cloning
  • EXPRESSION
    CONSTRUCT
    VERIFICATION
    • sequencing
    • Restriction digestion

Protein Expression

Aragen protein expression and purification services range from virtual protein sequences to the delivery of mg to g level proteins for compound screening assays, as well as for mechanistic and structural studies. Our solutions can be readily customized with a vast selection of vectors, promoters, affinity tags and protein labeling technologies, and our fast tracked methods enable us to generate functionally evaluated proteins in a compressed time frame of weeks.

We offer microbial, insect and mammalian expression systems for the generation of recombinant, functionally active proteins. Starting with database-led knowledge acquisition on the target protein, we select the appropriate expression system depending on target classification, post translational modifications and applications, to develop our workflow for target protein production.

Recombinant Protein Generation
Microbial
  • Soluble, inclusion bodies and periplasmic expression
  • Protein expression optimizations using vast E.coli expression strains
  • 50+ optimization parameters screening in 96 well format to Erlenmeyer flasks
  • Shake flask to fermenter scale-up
Insect Cell
  • Secretory and intracellular expression
  • Sf9, Sf21 and hi-five cell expression
  • Recombinant viral generation. Titration by conventional plaque assay/RT PCR approach
  • Protein expression optimization with MOI and PIH
  • Scale-up from shake flasks to WAVE bioreactors
Mammalian
  • Transient transfection
  • Intracellular/secretory/VLPL expressions
  • Lipofectamine and PEI based transfections
  • Adenoviral and lentiviral expression systems

Protein Purification & Characterization

With capabilities of purifying mg to g scale of proteins to >95% purity, and a throughput of 20 proteins per week using multiple ÄKTA purification platforms in a temperature controlled environment, we serve a wide range of pharmaceutical and agrochemical clients and academia. Our flexible, fully customizable protein purification services provide the highest purity and biological activity with low endotoxin levels. Having expressed over 200 reagent proteins, we consistently deliver superior quality proteins as reagents.

Recombinant Protein Generation
E.coli
BL21DE3, Rosetta, C41 and C43 expression hosts

  • Soluble
  • Inclusion bodies


Purification
  • Diverse column chemistry
  • IMAC, IEX, HIC, HPC, HAC, SEC and Affinity
  • Non-GLP/GMP production of pyrogen free sterile agent for cellular assays and animal studies



2-6 weeks of TAT

Insect
Sf9, Sf21 and high five expression hosts

  • Secretory protein expression
  • Soluble intracellular



QC
  • Purity determination by SDS PAGE
  • Western blot confirmation using target specific antibody of protein tagged antibody
  • Analytical SEC
  • LAL endotoxin assay (PTS/Kinetic QCL)
  • Tag removal
  • MALDI TOF and peptide fingerprinting; MS analysis

mg to gm level with 99% purity and endotoxin free

Mammalian
CHO, HEK, Expi293 and ExpiCHO expression hosts

  • Transient and stable expression
  • Secretory and soluble intracellular expression
  • Membrane protein expression and as VLPLS
Characterization
  • Activity assessment of the reagent with entire range of assay platforms
  • SPR on BIAcore T200
  • Western blot analysis
  • FACS
  • IEF (1D and 2D)
  • Custom based analysis

96-well format to column format of purification