PURIFICATION METHOD OF KHCV UB NS51.2 PROTEIN AS AN ANTIGEN SPECIFIC TO KOREAN HEPATITIS C VIRUS(HCV) FROM TRANSGENIC E. COLI

摘要

PURPOSE: Provided is a purification method of KHCV UB NS51.2 as an antigen specific to Korean hepatitis C virus(KHCV). Therefore, a more accurate HCV diagnosis method can be developed by using the purified UB NS5 1.2 proteins and further HCV vaccine can be developed by manufacturing a specific antibody to HCV antigen using the purified UB NS5 1.2 proteins. CONSTITUTION: The purification method comprises the steps of: a) obtaining insoluble precipitates by cell lysis of recombinant E. coli containing KHCV NS51.2 gene and a fused gene of ubiquitin gene (KHCV UB NS51.2 gene) and by centrifugation of the cell lysate; b) suspending the insoluble precipitates with a buffer solution containing triton X-100 then centrifuging the suspension to obtain insoluble precipitates; c) suspending the insoluble precipitates with a buffer solution containing a urea then centrifuging the suspension to obtain its supernatant; fractioning the supernatant by DEAE-sepharose ion exchange chromatography then collecting a fraction containing a KHCV UB 1.2 protein; fractioning the protein solution by 300 gel permeation chromatography to obtain a fraction containing KHCV UB NS5 1.2 protein; and dialyzing the collected protein solution to remove a urea and subjecting the resultant solution to FPLC-phenyl superose chromatography in order to purify KHCV UB NS5 1.2 protein.