Method for producing fusion protein having igg fc domain

Abstract

The present invention relates to a method for preparing a fusion protein having an IgG Fc domain and, specifically, to a method for preparing a fusion protein having an IgG Fc domain, the method additionally comprising a step of culturing cells, which produce the fusion protein, at a decreased culture temperature, thereby increasing cell growth and cell viability so as to increase fusion protein productivity and inhibiting aggregate generation so as to improve quality and production yield.

1 . A method for producing a fusion protein in a cell culture, comprising culturing cells at a decreased temperature of 28.0° C. or greater and less than or equal to 35.0° C. in order to increase an expression level of the protein, wherein the fusion protein comprises a soluble extracellular domain of a vascular endothelial growth factor (VEGF) receptor and a human immunoglobulin G (IgG) Fc domain. 
     
     
         2 . The method of  claim 1 , wherein the protein prepared by the method is a protein with a reduced amount of aggregates. 
     
     
         3 . The method of  claim 1 , wherein the cell culture is large-scale cell culture. 
     
     
         4 . The method of  claim 3 , wherein the cell culture is any one or more selected from a group consisting of batch culture, repeated batch culture, fed-batch culture, repeated fed-batch culture, continuous culture, and perfusion culture. 
     
     
         5 . The method of  claim 4 , wherein the cell culture is fed-batch cell culture. 
     
     
         6 . The method of  claim 1 , wherein the cell is a mammalian cell. 
     
     
         7 . The method of  claim 6 , wherein the mammalian cell is a CHO cell. 
     
     
         8 . The method of  claim 7 , wherein the CHO cell is a cell line of any one selected from a group consisting of DG44, DXB-11, K-1 and CHO-S. 
     
     
         9 . The method of  claim 1 , wherein a culture temperature from a culture initiation date until a temperature change is a temperature comprised in a temperature range of 33.0° C. to 38.0° C. 
     
     
         10 . The method of  claim 1 , wherein the decreased temperature is in a range of 30.0° C. to 35.0° C. 
     
     
         11 . The method of  claim 1 , wherein a culture period from a culture initiation date until a temperature change is 1 to 5 days. 
     
     
         12 . The method of  claim 1 , wherein a culture period after a temperature decrease is 2 to 15 days. 
     
     
         13 . The method of  claim 1 , wherein a sum of a culture period before the temperature change and a culture period after the temperature change is 3 days or more. 
     
     
         14 . The method of  claim 1 , wherein the soluble extracellular domain of the VEGF receptor comprises immunoglobulin-like domain 2 of a first VEGF receptor and immunoglobulin-like domain 3 of a second VEGF receptor. 
     
     
         15 . The method of  claim 1 , wherein the produced protein is a therapeutic protein. 
     
     
         16 . A method for preparing a target protein, the method comprising: culturing, through the method of  claim 1 , cells producing the target protein. 
     
     
         17 . The method of  claim 16 , further comprising a process of recovering the target protein from a culture broth in which the cells producing the target protein were cultured. 
     
     
         18 . The method of  claim 16 , wherein the target protein is a therapeutic protein. 
     
     
         19 . A pharmaceutical composition comprising the therapeutic protein prepared by the method of  claim 18  and a pharmaceutically acceptable carrier. 
     
     
         20 . The method of  claim 17 , wherein the target protein is a therapeutic protein. 
     
     
         21 . A pharmaceutical composition comprising a therapeutic protein prepared by the method of  claim 20  and a pharmaceutically acceptable carrier