E.coli Expression System

For low-expression cases, IPODIX Biotech prioritizes gene-host compatibility optimization via codon optimization or Rosetta strains to remedy rare codons. We systematically adjust induction conditions (OD, temperature, induction time, IPTG concentration). For host-toxic proteins, mild expression schemes or alternative expression platforms are applied. Sequencing and ORF checks are conducted to rule out construction defects.

For severely degraded proteins, all workflows are performed under low temperature with protease inhibitors supplemented in lysis and purification buffers. We reduce sample processing duration and speed up column purification. Strains with low protease background, revised tag positions or modified fusion schemes are adopted to improve protein stability as needed.

Solubility is improved via weakened induction (lower temperature, reduced IPTG, shorter induction time). Strains for soluble expression and MBP/GST/SUMO fusion tags are applied to aid folding. For heavily precipitated protein, inclusion body solubilization and refolding is adopted to harvest target protein.

Prokaryotic proteins are prone to endotoxin contamination that disturbs cell functional assays and animal experiments. We integrate endotoxin removal procedures after purification and refine workflows to prevent secondary pollution. Final proteins undergo endotoxin detection and control at IPODIX to meet various experimental demands.