Protein Case Studies: E. coli cell expression system

Listed below are several protein production case studies utilizing an E. coli expression system. For more details regarding our custom protein production services, click here. If you would like to browse the workflow utilized by our team in our E. coli protein production process, click here.

  • Protein requirement:

    • No tag, refolding of insoluble protein, high purity >98% by SEC-HPLC, endotoxin <50EU/mg

  • Strategy: 

    • Inclusion body solubilization

    • Refolding optimization 

    • No tag purification via IEX purification

    • Endotoxin removal: inclusion body washing, Q Sepharose FF column, endotoxin removal beads

  • Turnaround time: 6 weeks

  • Deliverables: 

    • 20mg no-tag protein

    • Purity >98%

    • Endotoxin <50EU/mg

Data

No tag, refolded and active cytokine mutant:

 

Human Heterodimer:

  • Protein requirements:

    • 50mg, endotoxin level < 1EU/ug> 90% SEC-HPLC purity

  • Strategy: 

    • His-tagged S100A8 (10.8kDa) and tag-free S100A9 (13.2kDa)

    • Dimer formation and purification by Ni-NTA column 

  • Deliverables: 

    • 50 mg S100A8/S100A9 heterodimer 

    • Purity >90% by reducing SDA-PAGE and SEC-HPLC

    • Endotoxin < 1EU/ug

Data

 
 

High-throughput production

  • Protein requirements:

    • Over 1000 non-commercially available antigen proteins

    • High yield, high purity, soluble proteins with no extra amino acid

    • 100 proteins/month

  • Deliverables: 

    • >2mg tag-free soluble proteins at 90% purity

    • Conc.>0.5mg/ml required for immunization

High-throughput production process