Genetically surface-modified Escherichia coli outer membrane vesicles targeting MUC1 antigen in cancer cells
Outer membrane vesicles (OMVs), non-replicating spherical liposomes derived from Gram-negative bacteria, are a promising vaccine platform and multifunctional delivery systems. Their ability to be modified via genetic engineering for the incorporation and display of heterologous proteins enhances the...
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Elsevier
2024-12-01
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| Series: | Biotechnology Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215017X24000274 |
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| author | Sedthawut Laotee Wanatchaporn Arunmanee |
| author_facet | Sedthawut Laotee Wanatchaporn Arunmanee |
| author_sort | Sedthawut Laotee |
| collection | DOAJ |
| description | Outer membrane vesicles (OMVs), non-replicating spherical liposomes derived from Gram-negative bacteria, are a promising vaccine platform and multifunctional delivery systems. Their ability to be modified via genetic engineering for the incorporation and display of heterologous proteins enhances their functionality. In this study, we demonstrated a bio-ligation approach to display single-chain variable fragments (scFv) on the OMV surface using the SpyTag/SpyCatcher system. SpyTag-fused scFv, expressed by mammalian cells, bound to OMVs with SpyCatcher-fused Lpp'OmpA after a simple incubation. Biophysical analysis indicated that the conjugated OMVs maintained their physicochemical properties. We used an scFv targeting mucin 1 protein (MUC1) for specific cell targeting. Confocal microscopy revealed that conjugated OMVs specifically bound to and were internalized by MUC1-presenting cells, but not by MUC1-deficient cells. In conclusion, this rapid and efficient bio-ligation system facilitates the display of functional scFv on OMV surfaces, offering a promising approach for targeted delivery to MUC1-expressing cancer cells. |
| format | Article |
| id | doaj-art-c4c51ac0e39242baadd1960f592d0dc8 |
| institution | Kabale University |
| issn | 2215-017X |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biotechnology Reports |
| spelling | doaj-art-c4c51ac0e39242baadd1960f592d0dc82024-12-06T05:13:38ZengElsevierBiotechnology Reports2215-017X2024-12-0144e00854Genetically surface-modified Escherichia coli outer membrane vesicles targeting MUC1 antigen in cancer cellsSedthawut Laotee0Wanatchaporn Arunmanee1Department of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, ThailandDepartment of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand; Center of Excellence in Cancer Cell and Molecular Biology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand; Corresponding author.Outer membrane vesicles (OMVs), non-replicating spherical liposomes derived from Gram-negative bacteria, are a promising vaccine platform and multifunctional delivery systems. Their ability to be modified via genetic engineering for the incorporation and display of heterologous proteins enhances their functionality. In this study, we demonstrated a bio-ligation approach to display single-chain variable fragments (scFv) on the OMV surface using the SpyTag/SpyCatcher system. SpyTag-fused scFv, expressed by mammalian cells, bound to OMVs with SpyCatcher-fused Lpp'OmpA after a simple incubation. Biophysical analysis indicated that the conjugated OMVs maintained their physicochemical properties. We used an scFv targeting mucin 1 protein (MUC1) for specific cell targeting. Confocal microscopy revealed that conjugated OMVs specifically bound to and were internalized by MUC1-presenting cells, but not by MUC1-deficient cells. In conclusion, this rapid and efficient bio-ligation system facilitates the display of functional scFv on OMV surfaces, offering a promising approach for targeted delivery to MUC1-expressing cancer cells.http://www.sciencedirect.com/science/article/pii/S2215017X24000274Outer membrane vesiclesSurface display systemSingle-chain variable fragmentSpyTag/SpyCatcher systemCell-specific delivery system |
| spellingShingle | Sedthawut Laotee Wanatchaporn Arunmanee Genetically surface-modified Escherichia coli outer membrane vesicles targeting MUC1 antigen in cancer cells Biotechnology Reports Outer membrane vesicles Surface display system Single-chain variable fragment SpyTag/SpyCatcher system Cell-specific delivery system |
| title | Genetically surface-modified Escherichia coli outer membrane vesicles targeting MUC1 antigen in cancer cells |
| title_full | Genetically surface-modified Escherichia coli outer membrane vesicles targeting MUC1 antigen in cancer cells |
| title_fullStr | Genetically surface-modified Escherichia coli outer membrane vesicles targeting MUC1 antigen in cancer cells |
| title_full_unstemmed | Genetically surface-modified Escherichia coli outer membrane vesicles targeting MUC1 antigen in cancer cells |
| title_short | Genetically surface-modified Escherichia coli outer membrane vesicles targeting MUC1 antigen in cancer cells |
| title_sort | genetically surface modified escherichia coli outer membrane vesicles targeting muc1 antigen in cancer cells |
| topic | Outer membrane vesicles Surface display system Single-chain variable fragment SpyTag/SpyCatcher system Cell-specific delivery system |
| url | http://www.sciencedirect.com/science/article/pii/S2215017X24000274 |
| work_keys_str_mv | AT sedthawutlaotee geneticallysurfacemodifiedescherichiacolioutermembranevesiclestargetingmuc1antigenincancercells AT wanatchapornarunmanee geneticallysurfacemodifiedescherichiacolioutermembranevesiclestargetingmuc1antigenincancercells |