Expression of secreted recombinant human insulin-like growth factor-II (IGF-II) in Chinese hamster ovary cells.
Chinese hamster ovary (CHO-KI) cells were cotransfected with a plasmid pcDNAI containing the human preproinsulin-like growth factor II cDNA linked downstream to the human cytomegalovirus promoter and with a plasmid containing the neomycin resistance gene (pMAM-neo). CHO neo+ were selected by growth in medium supplemented with G418 geneticin. After amplification, the neomycin-resistant clones were screened for IGF-II production. IGF-II produced was identified by dot blot and quantified by ELISA. The clones C24, C40 and C94 secreted IGF-II at about 350-400 ng per 10(6) cells per day. DNA analysis of C24 and C40 CHO cells by PCR demonstrated the presence of the IGF-II construct in the transfected cells, presumably integrated into the chromosomal DNA. IGF-II produced by CHO cells and purified by RP-HPLC was a mitogen for MCF-7 stimulating mitosis 2-fold.
Bekkari
H
H
Department of Biochemistry, Medicine School, University of Nancy, Vandoeuvre, France.
Sekkat
D
D
Straczek
J
J
Hess
K
K
Belleville-Nabet
F
F
Nabet
P
P
D000818:Animals; D001483:Base Sequence; D016466:CHO Cells / Q000378:metabolism; D006224:Cricetinae; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics; D006801:Humans; D007335:Insulin-Like Growth Factor II / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification
D018076:DNA, Complementary; D011994:Recombinant Proteins; D007335:Insulin-Like Growth Factor II