In vitro refolding process of bovine allergen β-lactoglobulin by Multispectroscopic method.
- Author:
Xu Li WU
1
;
Wen Pu WANG
;
Li Xin XIA
;
Hong XU
;
Hui WU
;
Zhi Gang LIU
Author Information
- Publication Type:Journal Article
- MeSH: Allergens; Animals; Cattle; Circular Dichroism; Enzyme-Linked Immunosorbent Assay; Immunoglobulin E; Lactoglobulins; chemistry; metabolism; Models, Molecular; Protein Binding; Protein Conformation; Protein Denaturation; Protein Folding; Spectrometry, Fluorescence; methods
- From: Biomedical and Environmental Sciences 2012;25(3):334-339
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo characterize the relationship between the refolding process of recombinant bovine β-lactoglobulin and its immunoreactivity for clinical purposes. To establish a spectral method which examine the extent of recombinant allergen renaturation.
METHODSThe refolding process of recombinant bovine β-lactoglobulin was investigated by using circular dichroism, fluorescence and synchronous fluorescence spectra. IgE-binding capacity of recombinant protein was analyzed by ELISA. In addition, bioinformatic methods were used to explain the spectral characteristics and analyze the relationship between the conformational changes and the immunoreactivity of the protein during renaturation in vitro.
RESULTSRenaturation of recombinant bovine β-lactoglobulin resulted in a more compact structure resembling the natural counterpart with stronger IgE-binding capacity.
CONCLUSIONThe degree of protein renaturation correlated with the IgE-binding capacity of the protein. Results from this study may be of help for food allergy therapy and development of vaccination in the future.
