Assessment of the Adsorption Activity of Bacterial Cells Using Yersinia pseudotuberculosis Model
https://doi.org/10.21055/0370-1069-2024-1-192-195
Abstract
Currently, the Russian Anti-Plague Institute “Microbe” produces diagnostic immunoglobulins used in laboratory diagnostics of plague pathogen. One of the important stages in obtaining this category of drugs is the process of adsorption and removal of cross-reacting antibodies to increase the specificity of the drug. For this purpose, inactivated cells of Yersinia pseudotuberculosis strains are used.
The aim of the work was to assess the possibility of using immunochemical methods to analyze the adsorption properties of bacterial cells, followed by an assessment of the impact of cultivation method and conditions, and the variant of the strain used on the adsorption properties of bacterial cells.
Materials and methods. Cultivation was carried out on solid and liquid nutrient media; cells were inactivated by temperature and chemical exposure. Adsorption activity was assessed using immunoblotting and inhibitory enzyme-linked immunosorbent assay. Statistical processing of the results was performed using two-factor analysis of variance.
Results and discussion. The suitability of immunoblotting and inhibitory ELISA methods for qualitative and quantitative assessment of the adsorption properties of bacterial cells has been demonstrated. It has been established that the adsorption properties of bacterial cells are influenced by the method of inactivation and the application of substrate feeding; the method of cultivation does not affect the adsorption properties of cells.
About the Authors
R. R. SalikhovRussian Federation
Ruslan R. Salikhov,
46, Universitetskaya St., Saratov, 410005
E. M. Kuznetsova
Russian Federation
46, Universitetskaya St., Saratov, 410005
O. A. Volokh
Russian Federation
46, Universitetskaya St., Saratov, 410005
References
1. Khushiramani R., Tuteja U., Shukla J., Batra H.V. Charac¬ terization of outer membrane proteins of Yersinia pestis and Yersinia pseudotuberculosis strains isolated from India. Indian J. Exp. Biol. 2004; 42(5):508–14.
2. Chen Y., Duan R., Li X., Li K., Liang J., Liu C., Qiu H., Xiao Y., Jing H., Wang X. Homology analysis and cross-immuno- genicity of OmpA from pathogenic Yersinia enterocolitica, Yersinia pseudotuberculosis and Yersinia pestis. Mol. Immunol. 2015; 68(2 Pt. A):290–9. DOI: 10.1016/j.molimm.2015.09.016.
3. Keasey S.L., Schmid K.E., Lee M.S., Meegan J., Tomas P., Minto M., Tikhonov A.P., Schweitzer B., Ulrich R.G. Extensive antibody cross-reactivity among infectious gram-negative bacteria revealed by proteome microarray analysis. Mol. Cell. Proteomics. 2009; 8(5):924–35. DOI: 10.1074/mcp.M800213-MCP200.
4. Sharma N., Hanif S., Upadhyay D., Chhikara M.K. Inhibition ELISA as a putative tool for the identification and quantification of meningococcal A and X polysaccharides at various stages of vac- cine development. J. Immunol. Methods. 2019; 473:112634. DOI: 10.1016/j.jim.2019.112634.
5. Salikhov R.R., Borisova S.V., Avdeeva N.G., Samokhvalova Yu.I., Volokh O.A. [Optimization of the conditions for cultivation of Yersinia pseudotuberculosis in the process of obtaining cell mass]. Problemy Osobo Opasnykh Infektsii [Problems of Particularly Dangerous Infections]. 2021; (4):137–42. DOI: 10.21055/0370-1069-2021-4-137-142.
6. Towbin H., Staehelin T., Gordon J. Electroforetic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl Acad. Sci. USA. 1979; 76(9):4350–4. DOI: 10.1073/pnas.76.9.4350.
7. Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680–5. DOI: 10.1038/227680a0.
8. Kobzar’ A.I. [Applied Mathematical Statistics. For Engineers and Scientists]. Moscow: “Fizmatlit”; 2006. 816 p.
Review
For citations:
Salikhov R.R., Kuznetsova E.M., Volokh O.A. Assessment of the Adsorption Activity of Bacterial Cells Using Yersinia pseudotuberculosis Model. Problems of Particularly Dangerous Infections. 2024;(1):192-195. (In Russ.) https://doi.org/10.21055/0370-1069-2024-1-192-195