Background: Romiplostim is a peptibody analogue of thrombopoietin (TPO) which regulates platelet production.\nThis molecule consists of two main parts: Peptide sequences which like wild type TPO, mimics stimulation of TPO\nreceptor and IgG1Fc, (Peptide + Antibody = Peptibody). This drug is used in treatment of chronic Immune\nThrombocytopenic Purpura (ITP).\nMethods: In this project E. coli bacteria were transformed by a construct harboring peptibody fusion gene. This\nconstruct consisted of two repeated peptide sequences which have fused to Carboxyl group of IgG1Fc. Designed\nconstruct in E. coli host resulted in protein expression in cytoplasm as inclusion body. The inclusion bodies were\nseparated, washed and after denaturation and solubilization, in the last stage the desired peptibodies were refolded\nand purified. The resulting peptibodies were characterized by SDS-PAGE and Western immunoblotting. The bioactivity\nwere assessed in vivo using subcutaneous injection in mice.\nResults: Results showed accurate molecules were produced and purified. Also, in vivo experiment showed significant\nincrement (more than two fold) of platelets compared to control group.\nConclusion: In this study laboratory scale production of recombinant romiplostim showed proper in-vivo bioactivity.\nThis new approach in expression and purification of this recently introduced thrombopoietin receptor agonist drug\nmay be followed by scale up of its production to response the chronic ITP patient�s demand.
Loading....