A method for the refolding or previously unfolded proteins with a concentrated solution of denaturing agent is presented, involving the use of high-performance hydrophobic interaction chromatography (H PHIC) to separate the denaturing agent completelyt from unfolded protein and to provide a suitable environment for its refolding The retention, peak shape and peak height in HPHIC and size-exclusion chromatography. UV spectra circular dichroic spectra and bioactivity were used to, test the possiblea and completeness of the protein refolding The proposed melhod permits the extracted solution from excherichia coli eells to be injected directly into to HPHIC column and, at the same time, the reflding and purification of the proteins to be effected the renaturation and purification of recombinant human interferon from Ecoli cells is one example of the application of the method in biotechnology