Experimental Substantiation of the Possibility to Use Finite Cell Line CHO-K1 for Determination of Specific Activity of Components of Chemical Cholera Vaccine
https://doi.org/10.21055/0370-1069-2019-4-113-116
Abstract
Objective was to experimentally substantiate the possibility to use the finite cell line CHO-K1 for measuring specific activity of cholera toxin and component of the vaccine choleragen-anatoxin in the process of chemical cholera vaccine manufacturing. Materials and methods. The studies involved the finite cell line CHO-K. The registration of results of bio-indication method was performed visually with the help of inverted microscope and photometrically - in colorimetric test for the assessment of metabolic activity of the cells at the wave length of 595 nm. Results and discussion. The proposed method allows for determining the toxin-production activity of Vibrio cholerae 569B strain during submerged cultivation in bioreactor and specific activity of choleragen-anatoxin by anatoxin binding measuring using cell cultures. The results correlate with the data obtained using intradermal Craig’s technique, GM1-ELISA and radial passive immune hemolysis (RPIH). Introduction of cell culture method into practice will provide for significant decrease in the volumes of usage of animals at the stages of manufacturing of chemical bivalent cholera vaccine.
About the Authors
O. S. DurakovaRussian Federation
Oksana S. Durakova.
46, Universitetskaya St., Saratov, 410005.
O. V. Gromova
Russian Federation
46, Universitetskaya St., Saratov, 410005.
A. V. Gaeva
Russian Federation
46, Universitetskaya St., Saratov, 410005.
S. V. Generalov
Russian Federation
46, Universitetskaya St., Saratov, 410005.
L. F. Livanova
Russian Federation
46, Universitetskaya St., Saratov, 410005.
O. D. Klokova
Russian Federation
46, Universitetskaya St., Saratov, 410005.
O. A. Volokh
Russian Federation
46, Universitetskaya St., Saratov, 410005.
References
1. Onishchenko G.G., Popova A.Yu., Kutyrev V.V., Smirnova N.I., Shcherbakova S.A., Moskvitina E.A., Titova S.V. [Relevant issues of epidemiological surveillance, laboratory diagnosis and prophylaxis of cholera in the Russian Federation]. Zhurnal Mikrobiologii, Epidemiologii i Immunobiologii [Journal of Microbiology, Epidemiology, and Immunobiology]. 2016; 1:89—101. DOI: 10.36233/0372-9311-2016-1-89-101.
2. Gaeva A.V., Gromova O.V, Durakova O.S., Generalov S.V, Volokh O.A. [Modern approaches to the control of active agents of chemical cholera vaccine*]. Razrabotka i Registratsiya Lekarstvennykh Sredstv [Development and Registration of Pharmaceuticals]. 2018; 1:152–7.
3. Craig J. Preparation of the vascular permeability factor of V cholera. J. Bacteriol. 1966; 92(3):793-5 PMID: 5922548. PMCID: PMC276327.
4. Shaginyan I.A., Marakusha B.M. [Modification of the method of passive immune hemolysis on solid nutrient medium for measuring the production of thermo labile enterotoxins by the strains of cholera vibrios and collibacillus]. Zhurnal Mikrobiologii, Epidemiologii i Immunobiologii [Journal of Microbiology, Epidemiology, and Immunobiology]. 1983; 2:92-6.
5. Ramamurthy T., Das B., Chakraborty S., Mukhopadhyay A.K., Sack D.A. Diagnostic techniques for rapid detection of Vibrio cholerae O1/O139. Vaccine. 2019; pii: S0264-410X(19)31020-5. DOI: 10.1016/j.vaccine.2019.07.099.
6. State Pharmacopeia of the Russian Federation, 13th edition. Moscow; 2015. Vol. 1-3.
7. Kumar P., Nagarajan A., Uchil P.D. Analysis of Cell Viability by the MTT Assay. Cold Spring Harb. Protoc. 2018; 6. DOI: 10.1101/pdb.prot095505.
8. Lakin G.F. [Biometry: Textbook for biology students of specialized higher educational institutions]. Moscow; 1990. 352p.
9. Guerrant R.L., Brunton L.L., Schaitman T.C., Rebhun L.I., Cilman A.C. Cyclic adenosine monophosphate and alteration of Chinese hamster ovary cell morphology: a rapid, sensitive in vitro assay for the enterotoxins of Vibrio cholerae and Escherichia coli. Infect. Immun. 1974; 10(2):320-7. PMID: 4368545. PMCID: PMC414999.
10. Freshni R.Ya. [Animal cell culture: Practice Guidelines]. Moscow; 2010. 714 p.
Review
For citations:
Durakova O.S., Gromova O.V., Gaeva A.V., Generalov S.V., Livanova L.F., Klokova O.D., Volokh O.A. Experimental Substantiation of the Possibility to Use Finite Cell Line CHO-K1 for Determination of Specific Activity of Components of Chemical Cholera Vaccine. Problems of Particularly Dangerous Infections. 2019;(4):113-116. (In Russ.) https://doi.org/10.21055/0370-1069-2019-4-113-116