E. coli is often the system of choice for biomanufacturing because of its amenability to genetic manipulation, low production cost, and rapid timelines for bioexpression.
Short affinity tags, such as polyhistidine and Strep-tag II, are often fused to recombinant proteins to aid in detection and purification of proteins.
1. Target gene(s) are cloned into LakePharma's proprietary bacterial expression plasmids with N or C-terminal His and/or Strep tags.
2. 1 liter expression in E. coli is performed at 18 oC or 37 oC with IPTG induction and the protein is purified by affinity chromatography using nickel resin or tandem affinity chromatography using nickel and streptactin resins. Elution fractions are analyzed by PAGE & Western blot and pooled based on purity metrics established with the client.
1) Catalog No.Custom includes a one-step affinity purification.
2) Catalog No.Custom includes tandem affinity purification (>90% purity is typically achieved but is not guaranteed).
3) Catalog No.Custom includes tandem affinity purification and an SEC polishing step.
Please note: expression may be subsequently scaled up to 10 liters or more to produce protein quantities needed by clients.
Options available: endotoxin removal, affinity tag removal by protease cleavage (inquire), protein titer quantification (HPLC), analytical protein characterization including aSEC (HPLC), C-IEF, thermal stability analysis, mass-spec, and protein formulation optimization.
DNA encoding target protein
1. Purified protein
2. Study report
3. Certificate of Analysis
Please note: plasmid DNA is not a deliverable.