Expression of rhEPO-L-Fc fusion protein and analysis of its bioactivity and pharmacokinetics.
- Author:
Qiang ZHU
1
;
Zhihua HUANG
;
Yuliang HUANG
;
Yang QIN
Author Information
1. Department of Biochemistry and Molecular Biology, College of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
- Publication Type:Journal Article
- MeSH:
Animals;
CHO Cells;
Cricetinae;
Cricetulus;
Erythropoietin;
chemistry;
genetics;
metabolism;
pharmacokinetics;
Female;
Humans;
Immunoglobulin Fc Fragments;
biosynthesis;
genetics;
Male;
Rats;
Rats, Sprague-Dawley;
Recombinant Fusion Proteins;
biosynthesis;
chemistry;
genetics;
pharmacokinetics;
Recombinant Proteins
- From:
Chinese Journal of Biotechnology
2008;24(11):1874-1879
- CountryChina
- Language:Chinese
-
Abstract:
To prolong serum half-life of human Erythropoietin for better efficacy, a new form of recombinant human erythropoietin (rhEpo-L-Fc) was generated by fusion of a full length human erythropoietin gene and the Fc fragment of human IgG1 with flexible linker sequence. The fusion gene rhEPO-L-Fc was constructed by PCR, then inserted into expression vector pOptiVEC-TOPO, and expressed in Chinese Hamster Ovary cells deficient in the DHFR enzyme(CHO-dhfr-). The chimeric protein was purified by Protein A affinity chromatography, showed expected molecular weight and demonstrated a similar bioactivity compared to that of the native recombinant human erythropoietin (rhEPO) in an EPO-dependent cell-based assay. In vivo pharmacokinetic studies showed that the rhEPO-L-Fc had an elimination half-life of 27 h. In vivo efficacy studies showed that a single dose administration of rhEPO-L-Fc in rats increased the reticulocyte number in the peripheral blood significantly. These results demonstrated that the new engineered rhEPO-L-Fc may become alternative therapeutic approach to extend the half-time of rhEPO to treat anemia.
