Genetically surface-modified Escherichia coli outer membrane vesicles targeting MUC1 antigen in cancer cells
Year: 2024
Type: article
Abstract: • E. coli OMVs were engineered to display scFv via the SpyTag/SpyCatcher system. • scFv targeting MUC1 (scFv SM3 ) was successfully conjugated onto the surface of bioengineered OMVs. • OMVs presenting scFv SM3 demonstrated specific binding to and internalization into MUC1-expressing cells, highlighting their potential as a cell-specific delivery system. 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.
Source: Biotechnology Reports
Authors: Sedthawut Laotee, Wanatchaporn Arunmanee
Institution: Chulalongkorn University
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,070
OpenAlex ID: W4402252495