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J Immunol Methods. 2008 Jun 1;335(1-2):116-20. doi: 10.1016/j.jim.2008.02.012. Epub 2008 Mar 11.

The biological activity of a recombinantly expressed (His)(6)-tagged peanut allergen (rAra h 1) is unaffected by endotoxin removal.

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1
Department of Agricultural Sciences, Faculty of Life Sciences, University of Copenhagen, Denmark. lbjj@life.ku.dk

Abstract

The application of recombinant (His)(6)-tagged proteins in cell culture assays is associated with problems due to lipopolysaccharide (LPS) contamination. LPS stimulates cells of the immune system, thereby masking antigen-specific activation of T cells. Due to the affinity of LPS for histidine it is associated with difficulties to remove LPS from recombinant (His)(6)-tagged proteins. Here we describe that the Triton X-114 phase separation method can be used to remove LPS from (His)(6)-tagged proteins and that the recombinant proteins retain their biological activity.

PMID:
18377922
DOI:
10.1016/j.jim.2008.02.012
[Indexed for MEDLINE]
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