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

Louise Bjerremann Jensen; Anna Maria Torp; Sven Bode Andersen; Per Stahl Skov; Lars K. Poulsen; Edward F. Knol; Els van Hoffen

doi:10.1016/j.jim.2008.02.012

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

Louise Bjerremann Jensen, Anna Maria Torp, Sven Bode Andersen, Per Stahl Skov, Lars K. Poulsen, Edward F. Knol, Els van Hoffen

University of Groningen

Research output: Contribution to journal › Article › Academic › peer-review

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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 (C) 2008 Elsevier B.V. All rights reserved.

Original language	English
Pages (from-to)	116-120
Number of pages	5
Journal	Journal of Immunological Methods
Volume	335
Issue number	1-2
DOIs	https://doi.org/10.1016/j.jim.2008.02.012
Publication status	Published - 1-Jun-2008

Keywords

recombinant protein
LPS
cell culture assays
NECROSIS-FACTOR-ALPHA
CELL
INTERLEUKIN-10
PROLIFERATION
SUPPRESSION

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

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 (C) 2008 Elsevier B.V. All rights reserved.",

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author = "Jensen, {Louise Bjerremann} and Torp, {Anna Maria} and Andersen, {Sven Bode} and Skov, {Per Stahl} and Poulsen, {Lars K.} and Knol, {Edward F.} and {van Hoffen}, Els",

year = "2008",

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doi = "10.1016/j.jim.2008.02.012",

language = "English",

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T1 - The biological activity of a recombinantly expressed (His)(6)-tagged peanut allergen (rAra h 1) is unaffected by endotoxin removal

AU - Jensen, Louise Bjerremann

AU - Torp, Anna Maria

AU - Andersen, Sven Bode

AU - Skov, Per Stahl

AU - Poulsen, Lars K.

AU - Knol, Edward F.

AU - van Hoffen, Els

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AB - 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 (C) 2008 Elsevier B.V. All rights reserved.

KW - recombinant protein

KW - LPS

KW - cell culture assays

KW - NECROSIS-FACTOR-ALPHA

KW - CELL

KW - INTERLEUKIN-10

KW - PROLIFERATION

KW - SUPPRESSION

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DO - 10.1016/j.jim.2008.02.012

M3 - Article

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JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

IS - 1-2

ER -

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

Abstract

Keywords

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