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Endotoxin-free protein production—ClearColi™ technology

Nature Methods volume 10, page 916 (2013)Cite this article

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Abstract

A new Escherichia coli strain with a genetically modified lipopolysaccharide (LPS) molecule has been created that enables protein expression without the endotoxin. Proteins expressed and purified from ClearColi™ BL21(DE3) cells will not trigger LPS-related immune response in human cells, making them ideal for immunogenicity, toxicity and cytokine assays.

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Figure 1: Comparison of apolipoprotein A-I (ApoA1) expression in ClearColi™ BL21(DE3) and Lucigen's E. Cloni® EXPRESS BL21(DE3) cells.
Figure 2: Comparison of Limulus amebocyte lysate (LAL) response in protein derived from BL21(DE3) cells and ClearColi™ BL21(DE3) cells after Ni-column purification.
Figure 3: Comparison of endotoxic response from protein derived from ClearColi™ BL21(DE3) and BL21(DE3) cells.

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Authors and Affiliations

  1. Research Center Borstel, Borstel, Germany

    Uwe Mamat & Kathleen Wilke

  2. University of Michigan, Ann Arbor, Michigan, USA

    Ronald W Woodard

  3. Research Corporation Technologies, Tucson, Arizona, USA

    Chad Souvignier

  4. Lucigen Corp., Middleton, Wisconsin, USA

    David Mead, Eric Steinmetz, Kristin Terry, Chelsea Kovacich, Andrew Zegers & Curtis Knox

Authors
  1. Uwe Mamat
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  2. Ronald W Woodard
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  3. Kathleen Wilke
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  4. Chad Souvignier
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  5. David Mead
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  6. Eric Steinmetz
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  7. Kristin Terry
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  8. Chelsea Kovacich
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  9. Andrew Zegers
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  10. Curtis Knox
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Corresponding author

Correspondence to Curtis Knox.

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Mamat, U., Woodard, R., Wilke, K. et al. Endotoxin-free protein production—ClearColi™ technology. Nat Methods 10, 916 (2013). https://doi.org/10.1038/nmeth.f.367

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