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Investigation on a Freeze-Drying Process for Long-Term Stability of mRNA-LNPs

2025· preprint· W7117657573 on OpenAlexfundno aff
MD Faizul Hussain Khan, Ayyappasamy Sudalaiyadum Perumal, Amine Kamen

Bibliographic record

VenuePreprints.org · 2025
Typepreprint
Language
FieldBiochemistry, Genetics and Molecular Biology
TopicProtein purification and stability
Canadian institutionsnot available
FundersCanadian Bee Research FundNatural Sciences and Engineering Research Council of CanadaMcGill University
KeywordsDispersityThermostabilityTrehaloseGelatinSugarProtein stabilityHistidine

Abstract

fetched live from OpenAlex

Background: Thermostability remains a key bottleneck for equitable access to mRNA–LNP vaccines, largely due to cold-chain requirements. Objectives and methods: Here, we optimized freeze-drying formulations by screening excipients (sugars, sugar-alcohols, and proteins) and buffers to preserve mRNA–LNP physicochemical (size, polydispersity index -PDI and encapsulation efficiency -EE) and fluorescence intensity-based, functional integrity assay (in vitro transfection) during long-term storage of up to 6 months. Results: In a preliminary screening study, different sugars (sucrose, trehalose), sugar alcohol (mannitol), protein (gelatin) and different buffers (Tris, PBS, histidine) were evaluated. The preliminary result showed that sucrose and trehalose, along with Tris and histidine buffers, had a positive effect on maintaining the physicochemical properties during freeze-drying, while mannitol, gelatin and PBS buffer had a negative effect. Based on these findings, the optimized formulations containing sucrose/Tris, sucrose/histidine, trehalose/Tris and trehalose/histidine were chosen, and a stability study was performed at −80, −20, 4, and 20 °C for six months. Conclusions: Overall, except for the samples maintained at 20 °C, no significant changes in the physicochemical quality of the freeze-dried mRNA-LNPs were observed over six months at −80, −20, and 4 °C. The in vitro stability study demonstrated stability at 4 °C for four months across all formulations, while a formulation with sucrose/Tris maintained satisfactory stability even at 20 °C for the same duration. The main results of this study demonstrate the feasibility of storing mRNA drug products as solid formulations at non-freezing temperatures (≤ 4 °C).

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

How this classification was reachedexpand

Full frame distilled prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.

metaresearch head score (Codex)0.003
metaresearch head score (Gemma)0.003
Version: codex-gemma-dda1882f352aValidation status: machine_predicted_unvalidated
Candidate categoriesMeta-epidemiology (narrow)
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: none
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.416
Threshold uncertainty score0.999

Codex and Gemma teacher scores by category

CategoryCodexGemma
Metaresearch0.0030.003
Meta-epidemiology (narrow)0.0010.001
Meta-epidemiology (broad)0.0010.001
Bibliometrics0.0000.000
Science and technology studies0.0000.001
Scholarly communication0.0000.000
Open science0.0010.001
Research integrity0.0010.001
Insufficient payload (model declined to judge)0.0000.000

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.177
GPT teacher head0.374
Teacher spread0.197 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one teacher head, not a consensus.

Study designBench or experimental
Domainnot available
GenreEmpirical

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

Quick stats

Citations0
Published2025
Admission routes1
Has abstractyes

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