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Prot."],"published-print":{"date-parts":[[2026,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    SARS-CoV-2 viral infection can be inhibited by blocking the interaction between the viral spike protein and the human receptor angiotensin-converting enzyme 2 (hACE2). The development of specific spike inhibitors using nanobodies, the antigen-binding region of llamas\u2019 antibodies, arose as a promising therapeutic method against SARS-CoV-2. However, one limitation of nanobodies is that they cannot be used directly in the human body due to their susceptibility to degradation. Bacteria-based delivery systems provide site-specific targeted action that can circumvent nanobody degradation. Here, we report the development of a genetically modified bacterium expressing anti-SARS-CoV-2 nanobodies that can inhibit the interaction between the hACE2 receptor and the receptor-binding domain (RBD) of the spike protein.\n                    <jats:italic>Lactococcus lactis<\/jats:italic>\n                    , a human symbiont probiotic bacterium, was selected to express nanobodies attached to their cell surface. Our data shows that FLAG-tagged anti-SARS-CoV-2 nanobodies were detected on the cell surface of recombinant\n                    <jats:italic>L. lactis<\/jats:italic>\n                    strains by flow cytometry and immunofluorescence without permeabilization. Furthermore, nanobodies are functional and can bind the RBD region from the spike protein in a dose-dependent manner. Inhibition of the hACE2-RBD interaction in cellular assays was quantified using a pseudotype lentivirus that mimics SARS-CoV-2 in an adaptation of the neutralization assay. Our results suggested that the recombinant bacteria can inhibit viral infectivity in more than 50% compared with a control without bacteria in a neutralization assay. These outcomes suggest that the engineered strain can be used in the future as a new therapeutic tool in COVID-19 prevention.\n                  <\/jats:p>","DOI":"10.1007\/s12602-025-10758-1","type":"journal-article","created":{"date-parts":[[2025,9,11]],"date-time":"2025-09-11T10:20:25Z","timestamp":1757586025000},"page":"4206-4221","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Construction of Synthetic Probiotic Bacteria for In Situ Delivery of Anti-SARS-CoV-2 Nanobodies"],"prefix":"10.1007","volume":"18","author":[{"given":"Carolina E.","family":"Portero","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claire","family":"Smith","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuxi","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M. Raquel","family":"March\u00e1n-Rivadeneira","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shiyong","family":"Wu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yong","family":"Han","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2025,9,11]]},"reference":[{"key":"10758_CR1","doi-asserted-by":"publisher","first-page":"565","DOI":"10.1016\/S0140-6736(20)30251-8","volume":"395","author":"R Lu","year":"2020","unstructured":"Lu R, Zhao X, Li J et al (2020) Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding. 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