Objective To 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.Methods The 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.Results Renaturation of recombinant bovine β-lactoglobulin resulted in a more compact structure resembling the natural counterpart with stronger IgE-binding capacity.Conclusion The 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. Objective To characterize the relationship between the refolding process of recombinant bovine beta-lactoglobulin and its immunoreactivity for a clinical purpose. To establish a spectral method which examine the extent of recombinant allergen renaturation. Methods The refolding process of recombinant bovine beta-lactoglobulin was investigated by using circular dichroism, fluorescence and synchronous fluorescence spectra. IgE-binding capacity of recombinant protein was analyzed by ELISA. 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 Results. Renaturation of recombinant bovine β-lactoglobulin resulted in a more compact structure resembling the natural counterpart with enhanced IgE-binding capacity. Confc The Degree of protein renaturation correlated with the IgE-binding capacity of the protein. Results from this study may be o f help for food allergy therapy and development of vaccination in the future.