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However, while giant unilamellar vesicles (GUVs) have become a popular artificial cell platform, the impact of fundamental design features remains obscure. Here, we assembled GUVs composed primarily of two phospholipids: POPC, a phosphatidylcholine widely employed in GUV preparation, and DOPC, a phosphatidylcholine with a lower melting temperature and higher fluidity. We found that the phospholipid mixture leads to an increase in average GUV size (compared to POPC only) without compromising GUV yield (in contrast to DOPC only). Moreover, we studied two artificial cell functions: enzyme-driven protein-membrane docking and protein expression. Artificial cells prepared from the phospholipid mixture exhibited enhanced relocation of proteins to the membrane and enhanced protein expression. Furthermore, our data allowed us to identify the limiting factor that underlies the observed differences in protein expression between membrane formulations: the efficiency of macromolecular encapsulation. Altogether, our study underscores the importance of membrane engineering as a powerful strategy for advancing the rational development of artificial cells with functional capabilities.<\/jats:p>","DOI":"10.1021\/acssynbio.6c00401","type":"journal-article","created":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T20:01:49Z","timestamp":1790539309000},"page":"4217-4228","source":"Crossref","is-referenced-by-count":0,"title":["Membrane Formulation Impacts Vesicle-Based Artificial Cell Functions"],"prefix":"10.1021","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8475-9820","authenticated-orcid":true,"given":"Jordi","family":"Ventura-Cobos","sequence":"first","affiliation":[{"name":"Universitat Polite\u0300cnica de Vale\u0300ncia, Universitat de Vale\u0300ncia , , Camino de Vera s\/n , ,","place":["Vale\u0300ncia, Spain, 46022"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6357-949X","authenticated-orcid":true,"given":"A\u0301ngela","family":"Morella\u0301-Aucejo","sequence":"additional","affiliation":[{"name":"Universitat Polite\u0300cnica de Vale\u0300ncia, Universitat de Vale\u0300ncia , , Camino de Vera s\/n , ,","place":["Vale\u0300ncia, Spain, 46022"]},{"name":"Instituto de Salud Carlos III , , ,","place":["Madrid, Spain, 28029"]},{"name":"Universitat Polite\u0300cnica de Vale\u0300ncia, Centro de Investigacio\u0301n Principe Felipe , , C\/ Eduardo Primo Yu\u0301fera 3 , ,","place":["Valencia, Spain, 46100"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4411-9145","authenticated-orcid":true,"given":"Bastiaan C.","family":"Buddingh\u2019","sequence":"additional","affiliation":[{"name":"Eindhoven University of Technology , , P.O. Box 513, ,","place":["Eindhoven, The Netherlands, 5600 MB"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7973-2404","authenticated-orcid":true,"given":"Jan C. M.","family":"van Hest","sequence":"additional","affiliation":[{"name":"Eindhoven University of Technology , , P.O. Box 513, ,","place":["Eindhoven, The Netherlands, 5600 MB"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5873-9674","authenticated-orcid":true,"given":"Ramo\u0301n","family":"Marti\u0301nez-Ma\u0301n\u0303ez","sequence":"additional","affiliation":[{"name":"Universitat Polite\u0300cnica de Vale\u0300ncia, Universitat de Vale\u0300ncia , , Camino de Vera s\/n , ,","place":["Vale\u0300ncia, Spain, 46022"]},{"name":"Instituto de Salud Carlos III , , ,","place":["Madrid, Spain, 28029"]},{"name":"Universitat Polite\u0300cnica de Vale\u0300ncia, Centro de Investigacio\u0301n Principe Felipe , , C\/ Eduardo Primo Yu\u0301fera 3 , ,","place":["Valencia, Spain, 46100"]},{"name":"Universitat Polite\u0300cnica de Vale\u0300ncia , , Camino de Vera s\/n , ,","place":["Valencia, Spain, 46022"]},{"name":"Universitat Polite\u0300cnica de Vale\u0300ncia, Instituto de Investigacio\u0301n Sanitaria La Fe (IIS La Fe) , , Av Fernando Abril Martorell 106 , ,","place":["Valencia, Spain, 46026"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0094-1893","authenticated-orcid":true,"given":"Loai K. E. A.","family":"Abdelmohsen","sequence":"additional","affiliation":[{"name":"Eindhoven University of Technology , , P.O. Box 513, ,","place":["Eindhoven, The Netherlands, 5600 MB"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6987-9506","authenticated-orcid":true,"given":"Antoni","family":"Llopis-Lorente","sequence":"additional","affiliation":[{"name":"Universitat Polite\u0300cnica de Vale\u0300ncia, Universitat de Vale\u0300ncia , , Camino de Vera s\/n , ,","place":["Vale\u0300ncia, Spain, 46022"]},{"name":"Instituto de Salud Carlos III , , ,","place":["Madrid, Spain, 28029"]},{"name":"Universitat Polite\u0300cnica de Vale\u0300ncia , , Camino de Vera s\/n , ,","place":["Valencia, Spain, 46022"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,27]]},"reference":[{"key":"2026101610062749200_cit1","doi-asserted-by":"publisher","first-page":"769","DOI":"10.1021\/acs.accounts.6b00512","article-title":"Artificial Cells: Synthetic Compartments with Life-like Functionality and Adaptivity","volume":"50","author":"Buddingh\u2019","year":"2017","journal-title":"Acc. 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C.\n          \n          Engineering Interactive Artificial Cells Ph.D. Thesis 1 (Research TU\/e\/Graduation TU\/e), Chemical Engineering and Chemistry; Technische Universiteit Eindhoven, 2020."}],"container-title":["ACS Synthetic Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acssynbio.6c00401","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/asbcd6\/article-pdf\/doi\/10.1021\/acssynbio.6c00401\/68669880\/sb6c00401.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/asbcd6\/article-pdf\/doi\/10.1021\/acssynbio.6c00401\/68669880\/sb6c00401.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T14:30:53Z","timestamp":1792161053000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/asbcd6\/article\/15\/10\/4217\/5439537\/Membrane-Formulation-Impacts-Vesicle-Based"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,27]]},"references-count":41,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2026,9,27]]},"published-print":{"date-parts":[[2026,10,16]]}},"URL":"https:\/\/doi.org\/10.1021\/acssynbio.6c00401","relation":{},"ISSN":["2161-5063"],"issn-type":[{"value":"2161-5063","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,27]]}}}