원문정보
초록
영어
The erythropoietin is 34-38kDa glycoprotein containing three N- and one O-glycosylation sites. It is known as a regulatory cytokine produced in adult kidney or fetal liver stimulating hematopoiesis 1). But its half-life is not so long thus it is necessary to enhance the stability of EPO in vivo. It is thought that fusion type of stable protein molecular linked EPO would increase in vivo half-life. In previous studies of other groups, an albumin was suggested as an ideal candidate for fusion protein to increase half-life and activity because it is the major protein and it has a long half-life in human blood stream2). We designed the human albumin gene fused N-terminal of EPO and inserted (GGSGG)4-repeated linker between albumin and EPO3). CHO-dhfr - was co-transfected with albumin-EPOs fusion genes and dhfr gene. Fused albumin-EPOs proteins were determined and quantified by Western blot and ELISA. Hematopoietic efficacy was estimated using EPO dependent cell line F-36E in vitro and normothaemic mice (B6D2F1) in vivo. And we also evaluated in vivo half-life of those proteins in SD-rat. As a result, albumin-EPOs showed better efficacy and longer half-life than the standard EPO.