RECOMBINANT PRODUCTION OF THE P60 ANTIGEN OF LISTERIA MONOCYTOGENES
Main Article Content
Authors
Abstract
Listeria monocytogenes is a significant foodborne pathogen characterized by an intracellular life cycle, pronounced adaptive mechanisms, and a broad spectrum of virulence factors. The aim of this study was to obtain the recombinant p60 antigen of L. monocytogenes from isolates circulating in the Republic of Kazakhstan. Using recombinant DNA technology, a producer strain Escherichia coli ArcticExpress(DE3)/Lmp60 was generated, enabling the expression of the protein with a molecular mass of 50.3 kDa. The specificity of the recombinant p60 antigen was confirmed by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. The obtained results highlight the diagnostic value of the p60 protein of L. monocytogenes as a target antigen for the development of highly specific ELISA-based test systems aimed at the rapid identification of L. monocytogenes in food products.
Keywords
Listeria monocytogenes, recombinant antigen, p60, ELISA, diagnostics, expression system
Article Details
References
Manyi-Loh C.E., Lues R. Listeria monocytogenes and listeriosis: the global enigma // Foods. – 2025. –Vol.14. – P. 1266. doi:10.3390/foods14071266
Zakir S., Abdo S., Bushra M., Hussein J. A. Current epidemiologic status and public health importance of listeriosis: a review // Journal of Veterinary Physiology and Pathology. – 2022. – Vol. 1(3). – P. 61–68. doi:10.58803/jvpp.v1i3.10
Koncurat A., Sukalic T. Listeriosis: characteristics, occurrence in domestic animals, public health significance, surveillance and control // Microorganisms. – 2024. – Vol. 12. – P. 2055. doi:10.3390/microorganisms12102055
Lopes-Luz L., Mendonça M., Bernardes Fogaça M., Kipnis A., Bhunia A.K., Bührer-Sékula S. Listeria monocytogenes: review of pathogenesis and virulence determinants-targeted immunological assays // Crit Rev Microbiol. – 2021. – Vol. 47. – P. 647-666. doi: 10.1080/1040841X.2021.1911930.
Radoshevich L., Cossart P. Listeria monocytogenes: towards a complete picture of its physiology and pathogenesis // Nat Rev Microbiol. – 2018. – Vol. 16. – P. 32-46. doi: 10.1038/nrmicro.2017.126.
Lee B.H., Cole S., Badel-Berchoux S., Guillier L., Felix B., Krezdorn N., Hébraud M., Bernardi T., Sultan I., Piveteau P. Biofilm Formation of Listeria monocytogenes Strains Under Food Processing Environments and Pan-Genome-Wide Association Study // Front Microbiol. – 2019. – Vol. 10. – Р. 2698. doi: 10.3389/fmicb.2019.02698.
Roberts B.N., Chakravarty D., Gardner J.C. 3rd, Ricke S.C., Donaldson J.R. Listeria monocytogenes Response to Anaerobic Environments // Pathogens. – 2020. – Vol. 9. – Р.210. doi: 10.3390/pathogens9030210.
Matle I., Mbatha K.R., Madoroba E. A review of Listeria monocytogenes from meat and meat products: Epidemiology, virulence factors, antimicrobial resistance and diagnosis // Onderstepoort J Vet Res. – 2020. – Vol. 87. – Р. 1-20. doi: 10.4102/ojvr.v87i1.1869.
Meireles D., Pombinho R., Cabanes D. Signals behind Listeria monocytogenes virulence mechanisms // Gut Microbes. – 2024. – Vol.16. – №1. – Р. 2369564. doi: 10.1080/19490976.2024.2369564.
Matsuda Y., Yamauchi H., Hara H. Activation of inflammasomes and mechanisms for intracellular recognition of Listeria monocytogenes // Microbiol Immunol. – 2023. – Vol. 67(10). – P. 429-437. doi: 10.1111/1348-0421.13091
Quereda J.J., Morón-García A., Palacios-Gorba C., Dessaux C., García-Del Portillo F., Pucciarelli M.G., Ortega A.D. Pathogenicity and virulence of Listeria monocytogenes: A trip from environmental to medical microbiology // Virulence. – 2021. – Vol. 12. – P. 2509-2545. doi: 10.1080/21505594.2021.1975526.
Lathrop A.L., Bailey N.B., Kim K.P., Bhunia A.K. Pathogen-specific antigen target for production of antibodies produced by comparative genomics // Antibody Technology Journal. – 2014. – Vol. 4. – P. 13-22. doi: 10.2147/ANTI.S54848.
Bubert A., Schubert P., Kohler S., Frank R., Goebel W. Synthetic peptides derived from the Listeria-monocytogenes P60-protein as antigens for the generation of polyclonal antibodies specific for secreted cell-free L-monocytogenes P60-proteins // Appl Environ Microbiol. – 1994. – Vol. 60(9). – P. 3120-3127.
Coutu J.V., Morissette C., D’Auria S., Lacroix M. Development of a highly specific sandwich ELISA for the detection of Listeria monocytogenes, an important foodborne pathogen // Microbiology Research International. – 2014. – Vol. 2(4). – Р. 46-52.
Yu K.Y., Noh Y., Chung M., Park H.J., Lee N., Youn M., Jung B.Y., Youn B.S. Use of monoclonal antibodies that recognize p60 for identification of Listeria monocytogenes // Clin Diagn Lab Immunol. – 2004. – Vol. 11(3). – P. 446-51. doi: 10.1128/CDLI.11.3.446-451.2004
Beauchamp S., D'Auria S., Pennacchio A., Lacroix M. A new competitive fluorescence immunoassay for detection of Listeria monocytogenes // Analytical Methods, Royal Society of Chemistry. – 2012. – Vol. 4. – P. 4187-92.
Wiktorczyk-Kapischke N., Skowron K., Wałecka-Zacharska E. Genomic and pathogenicity islands of Listeria monocytogenes-overview of selected aspects // Front Mol. Biosci. – 2023. – Vol. 14;10. – P. 1161486. doi: 10.3389/fmolb.2023.1161486
Yu M., Yang J., Guo M. Is the LysM domain of L. monocytogenes p60 protein suitable for engineering a protein with high peptidoglycan binding affinity? // Bioengineered. – 2016. – Vol. 7(6). – P. 406-410. doi: 10.1080/21655979.2016.1200772
Schmidt R.L., Filak H.C., Lemon J.D., Potter T.A., Lenz L.L. A LysM and SH3-domain containing region of the Listeria monocytogenes p60 protein stimulates accessory cells to promote activation of host NK cells // PLoS Pathog. – 2011. – Vol. 7(11). – P. 2368. doi: 10.1371/journal.ppat.1002368
Griffin M.E., Klupt S., Espinosa J., Hang H.C. Peptidoglycan NlpC/P60 peptidases in bacterial physiology and host interactions // Cell Chem. Biol. – 2023. – Vol. 18;30(5). – P. 436-456. doi: 10.1016/j.chembiol.2022.11.001
Lim H.Y., Oh Y.K., Kim, J.S., Lim, Y.K. Lee, Y.S and Yoon, B.S. Production of Monoclonal Antibody for Listeria spp. P60 Protein Based on iap Gene // Journal of Food Hygiene and Safety. – 2013. – Vol. 8. – P. 25-29.
Wang W., Liu L., Song S., Xu L., Zhu J., Kuang H. Gold nanoparticle-based paper sensor for multiple detection of 12 Listeria spp. by P60-mediated monoclonal antibody // Food and Agricultural Immunology. – 2017. – Vol. 28:2 – P. 274-287. doi: 10.1080/09540105.2016.1263986