Production and characterization of long-acting recombinant human albumin-EPO fusion protein expressed in CHO cell.
A long-lasting recombinant human albumin-linker-erythropoietin (EPO) is a human albumin gene fused to the N-terminal of EPO with a (GGSGG)(n)-repeated linker inserted between albumin and EPO. Albumin-EPO fusion genes were co-transfected with the dhfr gene. Albumin-EPO fusion protein has three kinds of sub-types (IALE, AD2LE, AD1LE). Albumin-EPO fusion protein was quantified with human EPO ELISA. The in vitro efficacy of albumin-EPO fusion protein was estimated using F-36E cell, and in vivo efficacy of albumin-EPO fusion protein was estimated using normocythemic mice (B6D2F1). We also determined the in vivo half-life in a Sprague-Dawley rat. A PLA program analysis result demonstrated that the albumin-EPO fusion protein IALE is about 7.8-fold more potent than rHuEPO in increasing the hematocrit of normal mice.
Joung
Chan-Hi
CH
Boryung Central Research Institute, Boryung Pharmaceutical Co. Ltd., Ansan 425-839, South Korea.
Shin
Ju-Yeop
JY
Koo
Jae-Kyung
JK
Lim
Jin J
JJ
Wang
Jin-Sang
JS
Lee
Song-Jae
SJ
Tan
Hyun-Kwang
HK
Kim
Sang-Lin
SL
Lim
Sang-Min
SM
D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D002452:Cell Count; D045744:Cell Line, Tumor; D002850:Chromatography, Gel; D003001:Cloning, Molecular / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D004921:Erythropoietin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006207:Half-Life; D006801:Humans; D051379:Mice; D016133:Polymerase Chain Reaction; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins; D012156:Reticulocytes; D012709:Serum Albumin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics
D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D012709:Serum Albumin; D004921:Erythropoietin