In-Use Stability and Device Compatibility Define Clinically Actionable Handling Limits for a GMP-Produced Attenuated Listeria monocytogenes Vaccine Expressing GUCY2C
Year: 2026
Type: article
Abstract: 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 × 109 CFU/bag) maintained CFU recovery through 6 h, whereas low-dose bags (3 × 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.
Source: Vaccines
Authors: Jagmohan Singh, Taranjot Johar, Vannessa Scully, Scott A. Waldman, Babar Bashir, Adam E. Snook
Language: English
Subfield: Biotechnology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,258
Awards: OCCULT COLORECTAL CANCER ELIMINATION BY GUCY2C-DIRECTED IMMUNOTHERAPY, Off-the-shelf CAR-T Therapy via Tumor-Selective Immuno-engagers, Neuroprotection by the GUCY2C gut-brain axis in Parkinson's disease, OCCULT COLORECTAL CANCER ELIMINATION BY GUCY2C-DIRECTED IMMUNOTHERAPY, G9000170100, Targeting visceral pain through intestinal neuropod cell GUCY2C signaling, Ligand-Induced Recruitment of CAR-T Cells to Colorectal Cancers
OpenAlex ID: W7162018501