Development of a method for detection and quantitative analysis of engeneered endolysin LysAm24-SMAP in biological samples

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Дәйексөз келтіру

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Аннотация

In recent years modified bacteriophage lysins are widely investigated for the purposes of antibacterial therapy development. Thus, effective and precise methods for the quantitative analysis of these enzymes are of high demand. The enzyme-linked immunosorbent assay (ELISA) method has been developed for the detection of recombinant modified endolysin LysAm24-SMAP in biological samples. The optimal parameters for protein detection were determined, particularly, the influence of salt and the composition of the buffer system for samples preparation was studied. The applicability of the immunodetection system of the genetically engineered endolysin LysAm24-SMAP in various biological samples with enzyme concentrations from 0.4 ng/ml was demonstrated. Also, the influence of matrix effects in animals’ organs and tissues homogenates samples, producer strain lysates and their individual components during the analysis was assessed and it was shown that 0.65 M NaCl addition in the ELISA buffer is crucial for achieving correct results and reduces non-specific interactions in the case of LysAm24-SMAP. The effectiveness of the developed system in the immunochemical control of the bacteriolytic enzyme was confirmed.

Толық мәтін

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Авторлар туралы

A. Klimova

Gamaleya National Research Centre for Epidemiology and Microbiology, Ministry of Health of the Russian Federation; Sechenov First Moscow State Medical University (Sechenov University), Ministry of Health of the Russian Federation

Email: northernnatalia@gmail.com
Ресей, Moscow, 123098; Moscow, 119991

I. Grigoriev

Gamaleya National Research Centre for Epidemiology and Microbiology, Ministry of Health of the Russian Federation

Email: northernnatalia@gmail.com
Ресей, Moscow, 123098

D. Vasina

Gamaleya National Research Centre for Epidemiology and Microbiology, Ministry of Health of the Russian Federation

Email: northernnatalia@gmail.com
Ресей, Moscow, 123098

M. Anurova

Sechenov First Moscow State Medical University (Sechenov University), Ministry of Health of the Russian Federation

Email: northernnatalia@gmail.com
Ресей, Moscow, 119991

V. Gushchin

Gamaleya National Research Centre for Epidemiology and Microbiology, Ministry of Health of the Russian Federation; Lomonosov Moscow State University

Email: northernnatalia@gmail.com
Ресей, Moscow, 123098; Moscow, 119991

N. Antonova

Gamaleya National Research Centre for Epidemiology and Microbiology, Ministry of Health of the Russian Federation

Хат алмасуға жауапты Автор.
Email: northernnatalia@gmail.com
Ресей, Moscow, 123098

Әдебиет тізімі

  1. Gerstmans H., Rodríguez-Rubio L., Lavigne R., Briers Y. // Biochem. Soc. Trans. 2016. V. 44. P. 123–128. https://doi.org/10.1042/BST20150192
  2. Love M.J., Bhandari D., Dobson R.C.J., Billington C. // Antibiotics (Basel). 2018. V. 7. № 1. 17. https://doi.org/10.3390/antibiotics7010017.
  3. Huemer M., Shambat S.M., Brugger S.D., Zinkernagel A.S. // EMBO Rep. 2020. e51034. https://doi.org/10.15252/embr.202051034
  4. Baquero F. // Int. Microbiol. 2021. V. 24. P. 499–506. https://doi.org/10.1007/s10123-021-00184-y
  5. Antonova N.P., Vasina D.V., Lendel A.M., Usachev E.V., Makarov V.V., Gintsburg A.L. et al. // Viruses. 2019. V. 11. № 3. https://doi.org/10.3390/v11030284
  6. Gutiérrez D., Briers Y. // Curr. Opin. Biotechnol. 2021. V. 68. P. 15–22. https://doi.org/10.1016/j.copbio.2020.08.014
  7. Fursov M.V., Abdrakhmanova R.O., Antonova N.P., Vasina D.V., Kolchanova A.D., Bashkina O.A. et al. // Viruses. 2020. V. 12. P. 545. https://doi.org/10.3390/v12050545
  8. Tabatabaei M.S., Ahmed M. // Methods Mol. Biol. 2022. 2508. P. 115–134. https://doi.org/10.1007/978-1-0716-2376-3_10
  9. Antonova N.P., Vasina D.V., Rubalsky E.O., Fursov M.V., Savinova A.S., Grigoriev I.V. et al. // Biomolecules. 2020. V. 10. P. 440. https://doi.org/10.3390/biom10030440
  10. Dawson R.M., Liu C.Q. // Drug Dev. Res. 2009. V. 70. P. 481–498.
  11. Vasina D.V., Antonova N.P., Grigoriev I.V., Yakimakha V.S., Lendel A.M., Nikiforova M.A. et al. // Front. Microbiol. 2021. V. 12. https://doi.org/10.3389/fmicb.2021.748718
  12. Arshinov I.R., Antonova N.P., Grigoriev I.V., Pochtovyi A.A., Tkachuk A.P., Gushchin V.A. et al. // Applied Biochemistry and Microbiology. 2022. V. 58. Suppl. 1. https://doi.org/10.1134/S0003683822100027
  13. Alves N.J. // Antib Ther. 2019. V. 2 P. 33–39. https://doi.org/10.1093/abt/tbz002
  14. Minas K., McEwan N.R., Newbold C.J., Scott K.P. // FEMS Microbiol. Lett. 2011. V. 325. P. 162–169. https://doi.org/10.1111/j.1574-6968.2011.02424.x
  15. Li G., Howard S.P. // Methods Mol. Biol. 2017. V. 1615. P. 143–149.
  16. Jun S.Y., Jung G.M., Yoon S.J., Youm S.Y., Han H.-Y., Lee J.-H. et al. // Clin Exp Pharmacol Physiol. 2016. V. 43. P. 1013–1016. https://doi.org/10.1111/1440-1681.12613
  17. Grishin A.V., Lavrova N.V., Lyashchuk A.M., Strukova N.V., Generalova M.S., Ryazanova A.V. et al. // Molecules. 2019. V. 24. https://doi.org/10.3390/molecules24101879
  18. Ross G.M. S., Filippini D., Nielen M.W.F., Salentijn G.I. // Anal. Chem. 2020. V. 92. P. 15587–15595. https://doi.org/10.1021/acs.analchem.0c03740
  19. Adhya S., Merril C. R., Biswas B. // Cold Spring Harb. Perspect Med. 2014. V. 4. https://doi.org/10.1101/cshperspect.a012518
  20. Höltje J.-V. // Arch. Microbiol. 1995. V. 164. P. 243–254. https://doi.org/10.1007/BF02529958
  21. Chen T., Rao, Y., Li J., Ren C., Tang D., Lin T. et al. // Int. J. Mol. Sci. 2020. V. 21. https://doi.org/10.3390/ijms21020501
  22. Callewaert L., Michiels C.W. // J. Biosci. 2010. V. 35. P. 127–160. https://doi.org/10.1007/s12038-010-0015-5
  23. Liu R., Meng Q., Dai Y., Zhang Y. // Chinese journal of biotechnology. V. 39. P. 4482–4496. https://doi.org/10.13345/j.cjb.230241
  24. Xu H., Lu J.R., Williams D.E. // J. Phys. Chem. B. 2006. V. 110. P. 1907–1914. https://doi.org/10.1021/jp0538161
  25. Generalova L.V., Grigoriev I.V., Vasina D.V., Tkachuk A.P., Kruzhkova I.S., Kolobukhina L.V. et al. // Bulletin of RSMU. 2022. V. 1. P. 14–21. https://doi.org/10.24075/brsmu.2022.005
  26. Gushchin V.A., Ogarkova D.A., Dolzhikova I.V., Zubkova O.V., Grigoriev I.V., Pochtovyi A.A. et al. // Front Immunol. 2022. V. 13. https://doi.org/10.3389/fimmu.2022.1023164
  27. Antonova N., Vasina D., Lendel A., Usachev E., Makarov V., Gintsburg A. et al. // Viruses. 2019. V. 11. https://doi.org/10.3390/v11030284
  28. Stiller J., Jasensky A.-K., Hennies M., Einspanier R., Kohn B. // J. Vet. Diagn. Invest. 2016. V. 3. P. 235–243. https://doi.org/10.1177/1040638716634397
  29. Biswas S., Saha M.K. // Immunochemistry & Immunopathology. 2015. V. 1. https://doi.org/10.4172/icoa.1000109

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