V.M. Popova1, A.A. Khakonov2, I.N. Matveeva3, V.T. Nochevny4, B.L. Manin5

1 FGBNU "All-Russian Research and Technological Institute of Biological Industry" (FGBNU "VNITIBP")
(Moscow Region, Losino-Petrovsky, Russia)

2 FGBI "All-Russian State Center for Quality and Standardization of Medicines for Animals and Feed" (FSBI "VGNKI") (Moscow, Russia)

3 FGBI "Federal Center for Animal Health Protection" (FGBI "ARRIAH") (Vladimir, Russia)

Improvement and development of modern technological processes for the production of culture models that meet the requirements of GMP for the manufacture of antiviral drugs.

Purpose of the work – development of modern technological processes for the production of virus-containing materials; search for new means, methods of control and production of high-quality cell cultures and active antigens in industrial volumes.

The success of the production and quality control of immunobiological preparations (UPS) consists in the search and certification of the most promising cell substrates, their availability and stability of biological properties, high sensitivity and absence of contamination, as well as the manufacturability of manufacturing and quality control of test cultures. An important place is occupied by the issues of effective control of nutrient media (PS) and blood serum (SC) of animals, selection and selection of the most productive cell models, as well as the use of the most effective laboratory and industrial methods of culturing viruses.

Long-term studies on the problem of optimizing the production of cell cultures made it possible to determine the issues of comparative assessment of the growth properties of commercial series of PS and STEM of animals and contamination by viruses, fungi and mycoplasmas. Antibacterial and antifungal drugs with a wide spectrum of action, as well as an alternative method of tissue disaggregation with a dispersant based on trisodium citrate (TNC), have been tested. Methods for obtaining and decontamination of animals with the help of lanthanides have been developed.

The practical application of the developed rules and requirements for the manufacture and control of culture models and starting materials can significantly expand the possibilities for standardizing production, increasing the activity of the final product, as well as increasing the efficiency and efficiency of the production of antiviral drugs.

Popova V.M., Khakonov A.A., Matveeva I.N., Nochevny V.T., Manin B.L. Improvement and standardization of technology for the production of cell cultures. Technologies of Living Systems. 2022. V. 19. № 3. Р. 63-72. DOI: https://doi.org/10.18127/j20700997-202203-07 (In Russian)

1. Zhivotnaya kletka v kulture (Metody i primeneniye v biotekhnologii.). Pod red. L.P. Diakonova. V.I. Sitkova. M.: Sputnik. 2000. 400 s. (in Russian).
2. Nochevnyy V.T., Kolyshkin V.M., Popov V.F. Perspektivy ispolzovaniya metoda protochnoy tsitometrii dlya standartizatsii perevivayemykh liniy kletok. Materialy mezhdunar. nauchn. konf. Vladimir. 2003. S. 498–500. (in Russian).
3. Avtorskoye svidetelstvo № 1067940. kl. S-12 № 9/64. 1984. Sposob polucheniya tripsina. V.T. Nochevnyy, D.F. Osidze, A.I. Guslavskiy, N.A. Bashashkina, A.G. Pochtarenko, G.E. Divarova, Yu.P. Chernetskiy, E.E. Shkolnikov, V.N. Kachalov. (in Russian).
4. Avtorskoye svidetelstvo № 1628526. SU 162852 A1 1990 g. Sposob polucheniya tripsina. V.T. Nochevnyy, V.F. Osidze, A.I. Guslavskiy. (in Russian).
5. Patent № 2403285 S (RU). 10.11. 2010. Opublikovano 10.11. 2010. Byul. № 31. C. 743. Zayavka № 2009121160/10 ot 04.06.2009. Sposob polucheniya tripsina. V.M. Popova, A.Ya. Samuylenko, A.I. Guslavskiy, I.L. Bero, E.E. Shkolnikov, A.A. Nezhuta, V.I. Eremets, V.A. Lukina, T.A. Skotnikova, L.A. Skorokhodova, N.K. Eremets, I.I. Kochish. (in Russian).
6. Patent № 2437936 CI (RU). 27.12.2011. Byul. № 36 Zayavka № 2010124270/10 ot 17.06.2010. Sposob immobilizatsii tripsina. V.M. Popova, V.A. Lukina, E.I. Yarygina, L.A. Skorokhodova, A.I. Guslavskiy, A.Ya. Samuylenko, I.I. Kochish, I.N. Matveyeva, V.I. Eremets, S.A. Grin, A.A. Rayevskiy, I.L. Bero, M.A. Frolova, N.K. Eremets. (in Russian).
7. Patent 2142503 (RF) 6.S.12 № 1/00.05/02. GNTsV i B «Vektor» Zayavl. 01.07.97. opubl. 10.12.99., Byul. 34. Sposob polucheniya sterilnogo rastvora tripsina dlya kultivirovaniya kletok. M.P. Bagryantseva, G.P. Troshkova, L.P. Kamshiy, L.D. Martynets, A.V. Velichko, V.T. Nochevnyy. (in Russian).
8. Nochevnyy V.T., Violin B.V., Karpukhina O.G. Enroflon-K – effektivnoye sredstvo profilaktiki mikoplazmennoy i bakterialnoy infektsii. Agrarnaya nauka. 2003. № 6. S 13–16. (in Russian).
9. Patent № 1365412 (RF). Preparat dlya predotvrashcheniya bakterialnoy kontaminatsii kultur tkaney. V.T. Nochevnyy, D.F. Osidze, V.F. Kovalev. (in Russian).
10. Petruchuk E.M., Nochevnyy V.T., Nikolayeva M.A. Perevivayemaya liniya kletok MDBK – kulturalnaya model dlya kontrolya syvorotki krovi na kontaminanty. Vestnik s-kh nauki. 1989. № 3. S. 72–77. (in Russian).
11. Bezgin V.M., Bukova N.K., Kozlov V.E. i dr. Osnovy promyshlennoy biotekhnologii. Kursk: Izd. KGSKhA. 2011. 512 s. (in Russian).
12. Kolokoltseva T.D., Saburina I.N. Patologicheskiye aspekty mikoplazmennoy kontaminatsii kletochnykh kultur. Patogenez. 2013. T. 11. № 3. S. 29–31. (in Russian).
13. Patent № 2460786 (KG) MPKS12N5/07. Sposob polucheniya syvorotki krovi vzroslogo krupnogo skota dlya kultivirovaniya kletok zhivotnykh i cheloveka. A.V. Ivanov, E.M. Plotnikova, N.I. Gurianov i dr. Zayavl. 19.04.2011 opubl. 10.09.2012. (in Russian).
14. Ponomarev A.P., Podkolzin I.V., Amelin V.G. Makro-, mikro- i ultramikroelementy v ekstraktakh iz prirodnogo nanotekhnologicheskogo minerala – shungita. Nanotekhnologii i okhrana zdorovia. 2012. T. IV. № 2. S. 48–55. (in Russian).
15. Patent № 2664729 (RF). MPK G01N 33/49/ Sposob ochistki syvorotki krovi krupnogo rogatogo skota ot kontaminiruyushchikh agentov. A.P. Ponomarev, E.V. Belik, B.L. Manin, M.M. Kogan. zayav. 23.05.17; opubl. 22.08.18. Byul. № 24. (in Russian).
16. Ponomarev A.P. Kultivirovaniye kletochnykh liniy cheloveka i zhivotnykh s ispolzovaniyem syvorotki krovi. ochishchennoy lantanoidami. Vladimir: Vlad. gos. un-t. 2019. S. 152–167. (in Russian).
17. Fedotov A.E. Osnovy GMP. M.: ASINKOM. 2012. 576 s. (in Russian).
18. Avtorskoye svidetelstvo 959422 (in Russian).
19. Nochevnyy V.T., Gabelko A.F. Kolichestvennyy metod biologicheskogo kontrolya pitatelnykh sred i syvorotki krovi zhivotnykh. Vestnik s.kh. nauk. 1992. № 2. S. 142–148. (in Russian).
20. Samuylenko A.Ya., Ruban E.A. Osnovy tekhnologii proizvodstva veterinarnykh biologicheskikh preparatov. M.: RASKhN. 2000. 781 s. (in Russian).
21. Laskovyy V.N., Nochevnyy V.T., Violin B.V. Formaldegid: metabolizm. antibakterialnyye. terapevticheskiye i immunomoduliruyushchiye svoystva. Agrarnaya nauka. 2005. № 10. S. 21–25. (in Russian).
22. Immunokorrektsiya v klinicheskoy veterinarnoy meditsine. Pod red. P.A. Krasochko Minsk: Tekhnoperspektiva. 2008. 507 s. (in Russian).
23. Glotov A.G., Shkilev N.A., Sergeyev A.N. Virusnyye i assotsiativnyye bolezni krupnogo rogatogo skota (Osobennosti epizootologii. patogeneza. klinicheskogo proyavleniya. patologicheskikh izmeneniy). Veterinarnyy konsultant. 2005. № 9. S. 5–14. (in Russian).