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Here, we leveraged retention using a selective hook (RUSH) system for the identification of human osteosarcoma U2OS cell secretion modulators, through automated, high-throughput screening of small compound libraries. We created a U2OS cell line which co-expresses a variant of streptavidin addressed to the lumen-facing membrane of the endoplasmic reticulum (ER) and a recombinant anti-PD-L1 antibody. The heavy chain of the antibody was modified at its C-terminus, to which a furin cleavage site, a green fluorescent protein (GFP), and a streptavidin binding peptide (SBP) were added. We show that the U2OS cell line stably expresses the streptavidin hook and the recombinant antibody bait, which is retained in the ER through the streptavidin\u2013SBP interaction. We further document that the addition of biotin to the culture medium triggers the antibody release from the ER, its trafficking through the Golgi where the GFP-SBP moiety is clipped off, and eventually its release in the extra cellular space, with specific antigen-binding properties. The use of this clone in screening campaigns led to the identification of lycorine as a secretion enhancer, and nigericin and tyrphostin AG-879 as secretion inhibitors. Altogether, our data support the utility of this approach for the identification of agents that could be used to improve recombinant production yields and also for a better understanding of the regulatory mechanism at work in the conventional secretion pathway.<\/jats:p>","DOI":"10.3390\/cells12121642","type":"journal-article","created":{"date-parts":[[2023,6,16]],"date-time":"2023-06-16T01:31:31Z","timestamp":1686879091000},"page":"1642","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Identification of Small Molecules Affecting the Secretion of Therapeutic Antibodies with the Retention Using Selective Hook (RUSH) System"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5187-0223","authenticated-orcid":false,"given":"Mathilde","family":"Coulet","sequence":"first","affiliation":[{"name":"Sanofi R&D, 94400 Vitry-sur-Seine, France"},{"name":"Metabolomics and Cell Biology Platforms, Gustave Roussy Cancer Center, 94800 Villejuif, France"},{"name":"Centre de Recherche des Cordeliers, Equipe Labellis\u00e9e par la Ligue Contre le Cancer, Universit\u00e9 de Paris Cit\u00e9, Sorbonne Universit\u00e9, Inserm U1138, Institut Universitaire de France, 75006 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sylvie","family":"Lachkar","sequence":"additional","affiliation":[{"name":"Metabolomics and Cell Biology Platforms, Gustave Roussy Cancer Center, 94800 Villejuif, France"},{"name":"Centre de Recherche des Cordeliers, Equipe Labellis\u00e9e par la Ligue Contre le Cancer, Universit\u00e9 de Paris Cit\u00e9, Sorbonne Universit\u00e9, Inserm U1138, Institut Universitaire de France, 75006 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marion","family":"Leduc","sequence":"additional","affiliation":[{"name":"Metabolomics and Cell Biology Platforms, Gustave Roussy Cancer Center, 94800 Villejuif, France"},{"name":"Centre de Recherche des Cordeliers, Equipe Labellis\u00e9e par la Ligue Contre le Cancer, Universit\u00e9 de Paris Cit\u00e9, Sorbonne Universit\u00e9, Inserm U1138, Institut Universitaire de France, 75006 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marc","family":"Trombe","sequence":"additional","affiliation":[{"name":"Sanofi R&D, 94400 Vitry-sur-Seine, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zelia","family":"Gouveia","sequence":"additional","affiliation":[{"name":"Institut Curie, PSL Research University, Sorbonne Universit\u00e9, CNRS, UMR 144, 26 rue d\u2019Ulm, 75005 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9129-9401","authenticated-orcid":false,"given":"Franck","family":"Perez","sequence":"additional","affiliation":[{"name":"Institut Curie, PSL Research University, Sorbonne Universit\u00e9, CNRS, UMR 144, 26 rue d\u2019Ulm, 75005 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6081-9558","authenticated-orcid":false,"given":"Oliver","family":"Kepp","sequence":"additional","affiliation":[{"name":"Metabolomics and Cell Biology Platforms, Gustave Roussy Cancer Center, 94800 Villejuif, France"},{"name":"Centre de Recherche des Cordeliers, Equipe Labellis\u00e9e par la Ligue Contre le Cancer, Universit\u00e9 de Paris Cit\u00e9, Sorbonne Universit\u00e9, Inserm U1138, Institut Universitaire de France, 75006 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9334-4405","authenticated-orcid":false,"given":"Guido","family":"Kroemer","sequence":"additional","affiliation":[{"name":"Metabolomics and Cell Biology Platforms, Gustave Roussy Cancer Center, 94800 Villejuif, France"},{"name":"Centre de Recherche des Cordeliers, Equipe Labellis\u00e9e par la Ligue Contre le Cancer, Universit\u00e9 de Paris Cit\u00e9, Sorbonne Universit\u00e9, Inserm U1138, Institut Universitaire de France, 75006 Paris, France"},{"name":"Department of Biology, Institut du Cancer Paris CARPEM, H\u00f4pital Europ\u00e9en Georges Pompidou, AP-HP, 75015 Paris, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"St\u00e9phane","family":"Basmaciogullari","sequence":"additional","affiliation":[{"name":"Sanofi R&D, 94400 Vitry-sur-Seine, France"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,6,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1007\/978-1-4939-3804-9_1","article-title":"ER to Golgi-Dependent Protein Secretion: The Conventional Pathway","volume":"1459","author":"Viotti","year":"2016","journal-title":"Methods Mol. 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