The Creation of Technological Lines for the Cultivation of Modified Lines of Mammalian Cells Expressing Recombinant Proteins
https://doi.org/10.35825/2587-5728-2017-1-1-33-41
EDN: yoiibn
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.
About the Authors
I. A. Churkin
Federal State Budgetary Establishment «48 Central Scientific Research Institute» of the Ministry of Defence of the Russian Federation
Russian Federation
Russian Federation
Churkin Igor Alekseevich. Chief of the Department. Candidate of Biological Sciences
Oktyabrskaya Street 11, Sergiev Posad 141306
S. V. Borisevich
Federal State Budgetary Establishment «48 Central Scientific Research Institute» of the Ministry of Defence of the Russian Federation
Russian Federation
Russian Federation
Chief. Doctor of Biological Sciences, Professor, Corr. Member RAS
Oktyabrskaya Street 11, Sergiev Posad 141306
D. A. Kutaev
Federal State Budgetary Establishment «48 Central Scientific Research Institute» of the Ministry of Defence of the Russian Federation
Russian Federation
Russian Federation
Deputy Chief for Scientific and Research Work. Candidate of Biological Sciences
Oktyabrskaya Street 11, Sergiev Posad 141306
V. T. Lymar
Federal State Budgetary Establishment «48 Central Scientific Research Institute» of the Ministry of Defence of the Russian Federation
Russian Federation
Russian Federation
Senior Researcher. Doctor of Medical Sciences
Oktyabrskaya Street 11, Sergiev Posad 141306
Yu. I. Pashchenko
Federal State Budgetary Establishment «48 Central Scientific Research Institute» of the Ministry of Defence of the Russian Federation
Russian Federation
Russian Federation
Leading Researcher. Doctor of Biological Sciences, Professor
Oktyabrskaya Street 11, Sergiev Posad 141306
E. V. Gordeev
Federal State Budgetary Establishment «48 Central Scientific Research Institute» of the Ministry of Defence of the Russian Federation
Russian Federation
Russian Federation
Deputy Chief of the Department
Oktyabrskaya Street 11, Sergiev Posad 141306
V. S. Kulish
Federal State Budgetary Establishment «48 Central Scientific Research Institute» of the Ministry of Defence of the Russian Federation
Russian Federation
Russian Federation
Deputy Chief of the Department. Candidate of Biological Sciences
Oktyabrskaya Street 11, Sergiev Posad 141306
T. M. Plekhanova
Federal State Budgetary Establishment «48 Central Scientific Research Institute» of the Ministry of Defence of the Russian Federation
Russian Federation
Russian Federation
Senior Researcher. Candidate of Biological Sciences
Oktyabrskaya Street 11, Sergiev Posad 141306
R. A. Khamitov
Federal State Budgetary Institution «State Scientific-Research Institute of Genetics and Plant Breeding of Industrial Microorganisms»
Russian Federation
Russian Federation
Vice Director for Strategic Development. Doctor of Medical Sciences, Professor
1 Dorozhny Drive 1, Moscow 117545
E. N. Sautkina
Federal State Budgetary Institution «State Scientific-Research Institute of Genetics and Plant Breeding of Industrial Microorganisms»
Russian Federation
Russian Federation
Chief of the Department of Medical Biotechnology. Candidate of Chemical Sciences
1 Dorozhny Drive 1, Moscow 117545
References
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Review
For citations:
Churkin I.A., Borisevich S.V., Kutaev D.A., Lymar V.T., Pashchenko Yu.I., Gordeev E.V., Kulish V.S., Plekhanova T.M., Khamitov R.A., Sautkina E.N. The Creation of Technological Lines for the Cultivation of Modified Lines of Mammalian Cells Expressing Recombinant Proteins. Journal of NBC Protection Corps. 2017;1(1):33-41. (In Russ.) https://doi.org/10.35825/2587-5728-2017-1-1-33-41. EDN: yoiibn
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