The biological activity of a recombinantly expressed (His)(6)-tagged peanut allergen (rAra h 1) is unaffected by endotoxin removal
Research output: Contribution to journal › Journal article › Research › peer-review
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.
| Original language | English |
|---|---|
| Journal | Journal of Immunological Methods |
| Volume | 335 |
| Issue number | 1-2 |
| Pages (from-to) | 116-20 |
| Number of pages | 5 |
| ISSN | 0022-1759 |
| DOIs | |
| Publication status | Published - 2008 |
- Allergens, Antigens, Plant, Basophils, Cells, Cultured, Cloning, Molecular, Dose-Response Relationship, Drug, Glycoproteins, Histamine Release, Histidine, Humans, Interleukin-10, Leukocytes, Mononuclear, Lipopolysaccharides, Plant Proteins, Polyethylene Glycols, Protein Binding, Recombinant Proteins, T-Lymphocytes, Time Factors, Tumor Necrosis Factor-alpha
Research areas
ID: 8109927