Related Experiment Video
Updated: Aug 27, 2025
06:28
Isolation of Low Endotoxin Content Extracellular Vesicles Derived from Cancer Cell Lines
Published on: February 17, 2023
1.2K
Plant produced endotoxin binding recombinant proteins effectively remove endotoxins from protein samples
Md Rezaul Islam Khan1, Muthamilselvan Thangarasu1, Hyangju Kang2
1Department of Life Sciences, Pohang University of Science and Technology, Pohang, 37673, Korea.
Scientific Reports
|September 30, 2022
Summary
A novel plant-produced fusion protein, CES3:CBM3, effectively removes toxic lipopolysaccharides (LPS) from recombinant proteins. This offers a cost-effective and robust platform for purifying biopharmaceuticals.
Area of Science:
- Biotechnology
- Protein Purification
- Biochemistry
Background:
- Lipopolysaccharides (LPS) are toxic contaminants in E. coli-produced recombinant proteins.
- Current LPS removal methods are challenging and expensive due to LPS solubility.
- Factor C's N-terminal domain (CES3) shows potential for LPS binding.
Purpose of the Study:
- To develop a plant-based platform for efficient lipopolysaccharide (LPS) removal from recombinant proteins.
- To investigate the use of horseshoe crab Factor C's N-terminal domain (CES3) for LPS binding and removal.
- To create a robust and cost-effective method for purifying biopharmaceuticals.
Main Methods:
- Expressed CES3 as part of a recombinant protein in Nicotiana benthamiana.
- Immobilized purified or microcrystalline cellulose (MCC) bead-bound CES3 for LPS binding assays.
- Generated Arabidopsis transgenic plants to produce CES3:CBM3 in an LPS-free environment.
- Utilized endogenous protease-mediated processing in plants to obtain functional CES3:CBM3:HDEL.
Main Results:
- Immobilized CES3 demonstrated strong binding to LPS-containing E. coli.
- Transgenic Arabidopsis plants produced a truncated, functional CES3:CBM3:HDEL protein.
- Plant-purified and MCC bead-immobilized CES3:CBM3:HDEL successfully removed LPS contamination.
- The CES3:CBM3 fusion protein proved effective in LPS remediation.
Conclusions:
- The CES3:CBM3 fusion protein produced in plants is a viable candidate for LPS removal.
- Immobilization onto MCC beads provides a practical and efficient platform for LPS decontamination.
- This plant-based system offers a promising, cost-effective solution for purifying recombinant proteins.
