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High-level expression and purification of the major house dust mite allergen Der p 2 in Escherichia coli.
- Source :
-
Protein Expression & Purification . May2016, Vol. 121, p97-102. 6p. - Publication Year :
- 2016
-
Abstract
- Der p 2, a major allergen derived from the house dust mite Dermatophagoides pteronyssinus , is one of the most clinically relevant allergens worldwide. Recombinant Der p 2 (rDer p 2) is useful in clinical diagnosis and disease-specific immunotherapy. However, previous studies showed that Der p 2 can only be expressed in Escherichia coli ( E. coli ) cells as inclusion bodies, thus protein refolding is required to obtain functional products. Here we report a new method to produce biologically active Der p 2 protein in E. coli . N-terminal hexahistidine- and trigger factor (TF)-tagged Der p 2 was expressed in soluble form in E. coli and purified using a combination of chromatography processes. This procedure produced milligram-level high purity Der p 2 per liter of bacterial culture. Moreover, far-UV region circular dichroism (CD) analysis and serum specific IgE reactivity test demonstrated that the secondary structure and IgE reactivity properties of rDer p 2 produced in our study were almost identical to those of natural Der p 2 (nDer p 2). In conclusion, the method developed in this work provides a useful tool for the production of immunologically active recombinant Der p 2 for clinical applications. [ABSTRACT FROM AUTHOR]
- Subjects :
- *HOUSE dust mites
*ALLERGENS
*ESCHERICHIA coli toxins
*AIR pollutants
*COLIFORMS
Subjects
Details
- Language :
- English
- ISSN :
- 10465928
- Volume :
- 121
- Database :
- Academic Search Index
- Journal :
- Protein Expression & Purification
- Publication Type :
- Academic Journal
- Accession number :
- 113896052
- Full Text :
- https://doi.org/10.1016/j.pep.2016.01.012