The biological activity of a recombinantly expressed (His)6-tagged peanut allergen (rAra h 1) is unaffected by endotoxin removal
- 作者：Louise Bjerremann Jensen ; Anna Maria Torp ; Sven Bode Andersen ; Per Stahl Skov ; Lars K. Poulsen ; Edward F. Knol ; Els van Hoffen
- 关键词：Recombinant protein ; LPS ; Cell culture assays
- 刊名：Journal of Immunological Methods
- 出版年：2008
- 期刊代码：57_00221759
- 类别：imm
- 出版时间：1 June 2008
- 卷：335
- 期：1-2
- 页码：116-120
- 文件大小：282 K
摘要
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.