OPTIMIZATION OF PRODUCTION OF RECOMBINANT ROMIPLOSTIM PEPTIBODY IN ESCHERICHIA COLI

Romiplastim is a fusion protein that has the same function as thrombopoietin in the body and thereby, stimulates bone marrow to produce more platelets. Thrombopoietin is a hormone that controls the production of platelets. Romiplastim is used in the treatment of thrombocytopenia in patients with chronic immune thrombocytopenia (ITP) when other drugs like corticosteroids and immunoglobulins or splenectomy have not treated them. The aim of this study was cloning and optimization of gene expression conditions and purification of recombinant Romiplastim peptibody in Escherichia coli bacteria.

In this practical study, the independent variables used included the amount of ODs (0.4, 0.8 and 1.2) at a wavelength of 600 nm, IPTG concentration (0.5, 1 and 1.5 mM) and the type of culture medium (LB, TB, M9). The Response Surface Methodology (RSM) was used in the form of central composite design with Design-Expert 12 software to predict independent variables on the amount of romiplastim production.

OD equal to 0.8, IPTG concentration equal to 1 mM and LB culture medium were optimized for the production of recombinant Romiplast peptibody.

Since the optimal production of peptibody was desired, cloning and optimization of conditions of gene expression and purification of Romiplastim peptibody in BL21 strain of Escherichia coli bacteria were done with the most optimal conditions which were done by RSM. Also, for correct folding of the protein, the refolding step was used at the beginning of the purification. Medicinal proteins play an important role in modern molecular medicine treatments.