[Baculovirus expression system-based expression of horseshoe crab factor C and its activity]
Lan Lan1, Huanlei Liu2,3,4,5,6, Hao Nan1,4
1College of Life Sciences, Northwest A & F University, Yangling 712100, Shaanxi, China.
Summary
This study presents a simplified, cost-effective method for producing recombinant factor C using a baculovirus system for sensitive endotoxin detection. This approach enhances yield and activity, paving the way for wider application in medical and pharmaceutical safety.
Area of Science:
- Biotechnology
- Molecular Biology
- Biochemistry
Context:
- Endotoxins pose significant risks in medical and pharmaceutical applications, necessitating reliable detection methods.
- The traditional Limulus amebocyte lysate (LAL) assay faces limitations including horseshoe crab depletion and assay variability.
- Recombinant factor C (rFC) offers a sensitive and quantifiable alternative but is hindered by high production costs and complex procedures.
Purpose:
- To develop a simplified and cost-effective method for producing recombinant factor C (rFC) using a baculovirus expression system.
- To enhance the yield and activity of rFC for improved endotoxin detection capabilities.
- To facilitate the widespread application of rFC by overcoming current production challenges.
Summary:
- A novel baculovirus expression system was engineered using a high-yield vector, optimal signal peptide, and codon optimization for rFC production.
- The method enables endotoxin detection directly from cell supernatant, significantly simplifying protein purification steps.
- The optimized system achieved a sensitivity of 0.05 EU/mL and supports 500,000 detection reactions per liter of fermentation broth.
Impact:
- This research streamlines rFC production, reducing costs and complexity for endotoxin detection.
- The enhanced yield and simplified purification process facilitate the broader adoption of rFC assays.
- Establishes a foundation for the increased use of rFC in ensuring safety in medical, pharmaceutical, and food industries.
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