TH35121C3 - Method for large-scale culture of human recombinant erythropoidin-producing cells. - Google Patents

Abstract

DC60 (08/10/55) The gene encoding human erythropoietin (EPO) was obtained from human genomic DNA. The gene contained sequences from the i) 5' region of the first transcribed ATG and ii) 3' of the stop codon of the EPO gene. The gene was cloned into an expression plasmid for cells containing true nuclei with an early SV40 viral promoter and promoter polyadinlation signaling as the only expression control fragment. Recombinant cells infected with the used genome resulted in unexpectedly high levels of recombinant EPO protein expression at a rate of 50 mg/L of culture medium per day. The gene encoding human erythropoietin (EPO) was obtained from human genomic DNA. The available gene consisted of sequences from the i) 5' region of the ATG that translates the first codon and ii) 3' region of the coton stop of the EPO gene. The EPO gene was cloned into an expression plasmid for cells containing true nuclei with an early SV 40 viral promoter, and promoter polyadinlation signaling is the only expression control fragment. Recombinant cells infected with the used gene resulted in unexpectedly high levels of protein expression of recombinant EPO at a dose of 50 mg/L of culture medium per day.

Claims (3)

All claims which will not appear on the advertisement page: 1. Where the nucleotide sequence excludes the non-coding fragments from the 5' and 3' regions, (b) the viral promoter, and (c) the viral terminator, under the conditions under which the polypeptide is expressed and recovered. 2. The method of claim 1 in which this condition is comprised of the exposure to methotrexate. 3. The method of claim 1 in which such culture produces more than 50 mg of EPO per liter of culture medium per day.