O-glycosylated alpha-interferon, used as medicament - isolated following secretion into conditioned medium of mammalian cells contg. a suitable expression plasmid

摘要

Isolation and purificn. of O-glycosylated interferon (IFN)-alpha from tissue culture supernatant of mammalian cells, pref. human embryonic kidney cells. These cells have been transformed with the IFN-alpha gene, or pref. the human IFN alpha or the human IFN alpha 2C gene, on a suitable expression plasmid, pref. pAD19B-IFN as claimed, and are grown under suitable condition. Also claimed is the produced IFN-alpha and its use as a medicament. USE - The IFN-alpha is used as a medicament (claimed) or as part of a medicament. In an example, the 5'-non-coding region of human IFN-alpha2C-encoding cDNA (derived from IF7; E. Dworkin-Rastl et al., J Interferon Res. 2 1982, 575-585) was replaced with the non-coding region of human beta-globin mRNA (Lawn et al., Cell 21, 1980, 647-651) using a polymerase chain reactors (PCR). A PCR product of 0.64 kilobases resulting from HindIII and XbaI restriction enzyme cuts was isolated and ligated into appropriate restriction enzyme sites of plasmid pAD-CMV19 (includes CMV promoter, modifier DHFR gene, splice donor and acceptor sites, SV40 early adenylating signal, SV40 originm, and multi-cloning site). E. coli HB101 were transformed with the ligation prods. and an expression plasmid pAD19B-IFN secreting O-glycosylated human IFN-alpha2C was identified.