Expression, Purification, and Immunological Characterization of Cr PI

Title: Expression, Purification, and Immunological Characterization of Cr PI

Volume: 14 Issue: 9

Author(s): Hai-Qiang Wu, Zhi-Gang Liu, Pi-Xin Ran, Zhen-Wen Zhou and Bo Gao

Affiliation:

Keywords: Periplaneta americana, Cr PI allergen, high-level expression, inclusion bodies, immunological characterization

Abstract: An efficient preparation of Periplaneta americana nymphae allergen, Cr PI (54 kDa) is described. It was expressed as a GST-tag fusion protein in Escherichia coli, strain BL21 (DE3). Expression of recombinant Cr PI (rCr PI), denaturation/ renaturation of the inclusion bodies and the effects of protein and L-arginine concentration on inclusion body aggregation were optimized. The fusion protein was purified by affinity chromatography and size exclusion chromatography, and Cr PI fusion protein was purified to > 95%. rCr PI bound strongly to IgE in the sera of individuals with cockroach allergies as shown by western blot and ELISA. Highly refolded and purified recombinant protein was obtained, providing a basis for the large-scale preparation of Cr PI allergen.

Cite this article as:

Wu Hai-Qiang, Liu Zhi-Gang, Ran Pi-Xin, Zhou Zhen-Wen and Gao Bo, Expression, Purification, and Immunological Characterization of Cr PI, Protein & Peptide Letters 2007; 14 (9) . https://dx.doi.org/10.2174/092986607782110266