E.Coli System Expression with Folding

Included in the package:

Steps in the process: Deliverables:
1. Cloning gene of interest- includes 1 optional  tag of choice (GST or 6XHis)
✓ 2 ug DNA
✓ Sequence chromatogram
2. Sub-cloning gene into expression vector
 
3. Expression, Folding, and Purification (>90%)
✓ 1 mg of protein
Reducing and Non-reducing SDS-PAGE image
4. QC analysis for endotoxin and mass spec ✓ Mass spec trace report
✓ LAL endotoxin report
5. QC conformational validation with 1 assay of choice (ELISA, WB, or Bioassay) ✓ 1 assay report
Additional options (additional charges via quotation):
Product manipulations (tag cleavage, desalting, reformulation, aliquoting, lyophilization, endotoxin removal) Add’l LAL endotoxin report
Additional quantitation method (Bradford, A280) Add’l quantitation report
Additional purified material Available in increments of 1 mg or more
Additional conformational validation assay  ✓ Add’l quality assay report
Time frame is 6-10 weeks for the package without additional options.

 

Expression Capabilities: Fermentation that scale to working volumes of 5L, 15L, and 65L

Purification Capabilities:

  • Ion Exchange (S-Sepharose, CM-Sepharose, Q-Sepharose, DEAE Sepharose)
  • Hydrophobic Interaction Chromatography (Butyl Sepharose, Phenyl Sepharose)
  • Hydroxyapatite Chromatography
  • Heparin Purification
  • Affinity Purification

 

Quality Control Capabilities

  • Mass spectroscopy – confirms MW of protein and can be used to determine multimeric status
  • Endotoxin analysis – Charles River PTS device measures endotoxin levels down to <0.05 EUs/ug
  • Western blotting - if antibodies are available
  • ELISA – if kits or antibodies are available
  • Bioassay -  See assay capabilities HERE