The Creation of Technological Lines for the Cultivation of Modified Lines of Mammalian Cells Expressing Recombinant Proteins

Abstract

The article presents data for creating technological line for the cultivation of modified lines of mammalian cells expressing recombinant proteins. This line is composed of three sections: for the creation of the modified lines of mammalian cells using modern molecular biological tools; for the implementation of scalable cultivation of genetically modified mammalian cells lines; for separation and purification of recombinant polypeptides using validated methods. Within the framework of the experimental study, the cells of СНО-S line, producing recombinant protein GP of the Ebola virus, have been grown up in the mixer reactor and in the wave-type fermenter with the use of suspension technology. It is established, that the wave-type fermenters have the advantage in the accumulation of cells and necessary protein. The recombinant polypeptide GP with purity exceeding 95 % has been isolated and purified from the cultural liquid. During the study of recombinant protein GP by ELISA and Western blot methods, it was detected that it's structure coincides with that of the native glycoprotein of the Ebola virus causative agent.

Keywords

• technological line
• cultivation
• mammalian cells
• expression
• recombinant protein