PURIFICATION METHOD OF RECOMBINANT HUMAN ERYTHROPOIETIN(EPO) EXPRESSED FROM RECOMBINANT INSECT CELLS

摘要

PURPOSE: A purification method of recombinant human erythropoietin(EPO) expressed from recombinant insect cells is provided, thereby rapidly and effectively purifying the human EPO, maintaining its activity. CONSTITUTION: A purification method of recombinant human erythropoietin(EPO) expressed from recombinant insect cells comprises the steps of: (1) culturing insect cells transformed with a microorganism containing an EPO gene; (2) subjecting the supernatant of the cultured medium to filtration, dialysis and centrifugation; (3) subjecting the concentrated medium to cation exchange chromatography to recover an EPO containing fraction; (4) subjecting the EPO containing fraction to lectin-affinity chromatography to isolate an EPO containing glycoprotein; and (5) subjecting the EPO containing glycoprotein to gel filtration chromatography, wherein sf-21 cell is infected with a human EPO containing baculovirus; the cation exchange chromatography uses a pH 6.0 to 6.5 buffer solution; the cation exchange chromatography uses CM-sepharose, S-sepharose, CM-cellulose, S-cellulose or mono-s resin; the lectin-affinity column uses ConA sepharose; and the gel filtration chromatography uses superose 12, superdex 75, sephadex 75 or sephacryl S-100.