{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T16:53:34Z","timestamp":1785430414521,"version":"3.56.0"},"reference-count":32,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T00:00:00Z","timestamp":1779321600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000005","name":"DoD","doi-asserted-by":"publisher","award":["W81XWH-22-1-0208"],"award-info":[{"award-number":["W81XWH-22-1-0208"]}],"id":[{"id":"10.13039\/100000005","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000005","name":"DoD","doi-asserted-by":"publisher","award":["W81XWH-22-1-0207"],"award-info":[{"award-number":["W81XWH-22-1-0207"]}],"id":[{"id":"10.13039\/100000005","id-type":"DOI","asserted-by":"publisher"}]},{"award":["W81XWH-22-1-0208"],"award-info":[{"award-number":["W81XWH-22-1-0208"]}],"id":[{"id":"https:\/\/ror.org\/042tbn311","id-type":"ROR","asserted-by":"publisher"}]},{"award":["W81XWH-22-1-0207"],"award-info":[{"award-number":["W81XWH-22-1-0207"]}],"id":[{"id":"https:\/\/ror.org\/042tbn311","id-type":"ROR","asserted-by":"publisher"}]},{"name":"NIH","award":["1R21 CA267087"],"award-info":[{"award-number":["1R21 CA267087"]}]},{"name":"NIH","award":["1R21 CA286339"],"award-info":[{"award-number":["1R21 CA286339"]}]},{"award":["1R21 CA267087"],"award-info":[{"award-number":["1R21 CA267087"]}],"id":[{"id":"https:\/\/ror.org\/044zkrt47","id-type":"ROR","asserted-by":"publisher"}]},{"award":["1R21 CA286339"],"award-info":[{"award-number":["1R21 CA286339"]}],"id":[{"id":"https:\/\/ror.org\/044zkrt47","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100012394","name":"Kleberg Foundation","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100012394","id-type":"DOI","asserted-by":"crossref"}]},{"id":[{"id":"https:\/\/ror.org\/05r5zfp34","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Targeted Diagnostic and Therapeutics, Inc"},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["P30 CA056036"],"award-info":[{"award-number":["P30 CA056036"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"award":["P30 CA056036"],"award-info":[{"award-number":["P30 CA056036"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Background: Live-attenuated Listeria monocytogenes (Lm) vectors are a clinically validated cancer immunotherapy platform, but translation requires reproducible, clinically realistic workflows for dose preparation and infusion. For live bacterial products, in-use stability and device compatibility can drive dose variability through adsorption, settling, and device losses. Methods: We developed and GMP-manufactured an attenuated Lm vaccine expressing human GUCY2C (Lm-GUCY2C) and performed translational characterization, including construct verification and immunogenicity readouts, and defined the administration-focused in-use stability and device compatibility. Post-thaw stability was assessed in primary cryovials and during preparation and delivery from 250 mL saline infusion bags using standard clinical devices (syringes\/needles, filter-free IV tubing) and OnGuard2 closed-system components. Samples were collected over 24 h at room temperature, and viable Lm-GUCY2C were quantified by CFU recovery. Results: Lm-GUCY2C remained stable in thawed cryovials for 24 h with no significant CFU loss. High-dose infusion bags (3 \u00d7 109 CFU\/bag) maintained CFU recovery through 6 h, whereas low-dose bags (3 \u00d7 108 CFU\/bag) exhibited significant losses beginning at 3 h, supporting a practical in-use window of up to 2 h for low-dose preparations. OnGuard2 intravenous (i.v.) connectors did not measurably affect CFU recovery, while OnGuard2 vial adapters reduced recovery. Conclusions: This work provides an end-to-end, translationally focused characterization of a GMP-manufactured Lm cancer vaccine, including clinically actionable in-use handling constraints and device compatibility. These data define preparation and administration guardrails (notably, time-to-infusion limits for low-dose bag preparations) that can improve dose accuracy and reproducibility in clinical testing.<\/jats:p>","DOI":"10.3390\/vaccines14050461","type":"journal-article","created":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T12:06:07Z","timestamp":1779365167000},"page":"461","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["In-Use Stability and Device Compatibility Define Clinically Actionable Handling Limits for a GMP-Produced Attenuated Listeria monocytogenes Vaccine Expressing GUCY2C"],"prefix":"10.3390","volume":"14","author":[{"given":"Jagmohan","family":"Singh","sequence":"first","affiliation":[{"name":"Department of Pharmacology, Physiology, and Cancer Biology, Thomas Jefferson University, Philadelphia, PA 19107, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taranjot","family":"Johar","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Physiology, and Cancer Biology, Thomas Jefferson University, Philadelphia, PA 19107, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vannessa","family":"Scully","sequence":"additional","affiliation":[{"name":"Investigational Drug Service, Thomas Jefferson University Hospital, Philadelphia, PA 19107, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6619-175X","authenticated-orcid":false,"given":"Scott A.","family":"Waldman","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Physiology, and Cancer Biology, Thomas Jefferson University, Philadelphia, PA 19107, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6843-1179","authenticated-orcid":false,"given":"Babar","family":"Bashir","sequence":"additional","affiliation":[{"name":"Department of Medical Oncology, Thomas Jefferson University, Philadelphia, PA 19107, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9216-4560","authenticated-orcid":false,"given":"Adam E.","family":"Snook","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Physiology, and Cancer Biology, Thomas Jefferson University, Philadelphia, PA 19107, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1038\/s41420-025-02652-0","article-title":"Development of a bispecific CDH17-GUCY2C ADC bearing the ferroptosis inducer RSL3 for the treatment of colorectal cancer","volume":"11","author":"Zhang","year":"2025","journal-title":"Cell Death Discov."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1007\/s00280-023-04507-w","article-title":"A phase I, first-in-human study of TAK-164, an antibody-drug conjugate, in patients with advanced gastrointestinal cancers expressing guanylyl cyclase C","volume":"91","author":"Kim","year":"2023","journal-title":"Cancer Chemother. 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