Patent ID: 6376218
Filing Date: 2002-04-23
Classification: C07K

Abstract:
A method of producing substantially purified recombinant human erythropoietin, comprising the steps of:culturing a baby hamster kidney fibroblast cell line expressing active recombinant human erythropoietin from an expression vector comprising a cDNA fragment encoding human erythropoietin, a plasmid vector pcDNA3.1 and a cytomegalovirus promoter, wherein said cDNA fragment is produced by polymerase chain reaction (PCR) using a primer set comprised of SEQ ID No. 1 and SEQ ID No. 2; collecting conditioned medium containing said expressed recombinant human erythropoietin from a culture of said baby hamster kidney fibroblast cell line; precipitating the human erythropoietin from said conditioned medium; applying a resuspension of precipitated human erythropoietin to an immobilized lectin column; eluting said human erythropoietin from said immobilized lectin column with an increased mannose gradient solution; applying eluted human erythropoietin to a gel filtration column; and, eluting the human erythropoietin from said gel filtration column.