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503O8,12
Formulation & Performance
Formulation & Performance
Study-specific compositions, preparation conditions and experimental outcomes for 503O8,12.
503O8,12-specific screening and spleen-biased expression reviewed against the main article. Previously reviewed SI characterization is retained as graph estimates, not exact tabulated measurements. Detailed lung cell-type and toxicity experiments belong to 514O6,10 and are excluded from this product’s claims.
1. Formulations and component amounts
Reported composition and calculated mass are separate. Molecular-weight inputs use catalog values or an explicitly identified reference average; DMG-PEG2000 is fixed initially at 2507. Edit inputs only when using a verified alternative material.
503O8,12 — 16PE screening / intravenous reporter formulation
Molar composition. Lipidoid:mRNA mass ratio 10:1. Reference-average PEG molecular weight 2693.32 is used for the calculation; batch-specific PEG distribution was not measured here.Main Results 2.2 and Experimental Section
| Component | Molecular weight | Molar parts | Normalized mol% | Amount (µmol) | Weight (mg) |
|---|---|---|---|---|---|
| 503O8,12 | 35.000 | 37.8088 | 60.4445 | ||
| DOPE | 16.000 | 17.2840 | 12.8598 | ||
| Cholesterol | 46.500 | 50.2316 | 19.4221 | ||
| 14:0 PEG2000 PE | 2.500 | 2.7006 | 7.2736 |
Calculated total: 100.0000 mg lipid. RNA, solvent and process loss are excluded.
503O8,12 — 40PE characterization variant
Molar composition; SI physical-characterization variant. Do not attach 514O6,10 lung transfection results to this composition.Main formulation method; previously reviewed SI Fig. S1
| Component | Molecular weight | Molar parts | Normalized mol% | Amount (µmol) | Weight (mg) |
|---|---|---|---|---|---|
| 503O8,12 | 35.000 | 34.6027 | 55.3189 | ||
| DOPE | 40.000 | 39.5459 | 29.4233 | ||
| Cholesterol | 22.500 | 22.2446 | 8.6009 | ||
| 14:0 PEG2000 PE | 2.500 | 2.4716 | 6.6569 |
Calculated total: 100.0000 mg lipid. RNA, solvent and process loss are excluded.
503O8,12 — 40TAP characterization variant
Molar composition; mass recipe withheld pending verified DOTAP salt / molecular-weight basis.Main formulation method; previously reviewed SI Fig. S1
| Component | Molecular weight | Reported molar entry | Calculated mass |
|---|---|---|---|
| 503O8,12 | 1598.69 | 35 | Not calculated |
| DOTAP (study salt basis unresolved) | Not confirmed | 40 | Not calculated |
| Cholesterol | 386.65 | 22.5 | Not calculated |
| 14:0 PEG2000 PE | 2693.32 | 2.5 | Not calculated |
No complete mass recipe is generated for this formulation while composition or identity-dependent inputs remain unresolved.
The study PEG-lipid is 14:0 PEG2000 PE. The reference average 2693.32 is used rather than the catalog DMG-PEG2000 value 2507. DOTAP identity-dependent input remains unresolved.
How component weights are calculated
For total lipid mass M (mg), molar parts rᵢ and molecular weight MWᵢ: component mass = M × rᵢ × MWᵢ / Σ(r × MW). Component amount (µmol) = component mass (mg) × 1000 / MWᵢ. Molar parts are normalized only for the displayed percentage. The calculation does not infer N/P or RNA dose.
2. Preparation and experimental setup
| Parameter | Reported procedure / limitation | Source |
|---|---|---|
| RNA / lipid relationship | Ionizable lipidoid:mRNA 10:1 by mass; reported mRNA formulation concentration 0.09 mg/mL. N/P is not supplied. | Experimental Section |
| Solvent and buffer | Lipid phase ethanol:citrate 9:1 by volume; RNA in 10 mM citrate pH 4. | Experimental Section |
| Mixing | Equal volumes of lipid and RNA phases; vortex 3,200 rpm for 5 seconds. | Experimental Section |
| Animal preparation | PBS dialysis using 3.5 kDa MWCO membrane for 1 h. | Experimental Section |
| Cargo | 5-methoxyuridine-modified reporter mRNA; polyU used in the initial TNS library screen. | Experimental Section; Results 2.2 |
| Mouse reporter assay | Female C57BL/6 mice, 6–8 weeks; 0.75 mg/kg mLuc by tail-vein injection. Luciferin at 3 h; tissue harvest approximately 15 min later. | Experimental Section; Figs. 2–3 |
| Cell reporter screen | HeLa cells, 15,000 per well; mLuc 100 ng/well, 24 h; n=4. | Experimental Section; Fig. 2c |
3. Particle characterization
| Measurement / batch | Reported result | Evidence context |
|---|---|---|
| 16PE diameter / encapsulation / zeta | Approximately 110 nm / 86% / −2.2 mV | Previously reviewed SI Fig. S1; visual graph estimates, n=4; no exact source-data values recovered |
| 40PE diameter / encapsulation / zeta | Approximately 100 nm / 88% / −4.8 mV | Previously reviewed SI Fig. S1; visual graph estimates, n=4 |
| 40TAP diameter / encapsulation / zeta | Approximately 105 nm / 83% / −0.6 mV | Previously reviewed SI Fig. S1; visual graph estimates, n=4 |
| PDI / apparent pKa / N/P | Not established for this product in the reviewed record | A TNS measurement at pH 5 does not determine pKa |
| Particle sizing method | Source method / caption discrepancy retained | Main methods describe nanoparticle tracking; SI caption describes a DLS number mean. Values are called mean particle size without resolving that inconsistency. |
4. Performance by experiment
| Experiment and dose | Readout and comparator | Result and source |
|---|---|---|
| Initial ionization screen PolyU-containing LNP library; TNS fluorescence at pH 5; n=12 | Readout: Plate-reader fluorescence Comparator: 580-member combinatorial library | 503O8,12 proceeded into reporter delivery evaluation. A single-pH screen is neither a pKa curve nor proof of in vivo delivery.Fig. 2a–c |
| Intravenous reporter potency 503O8,12 16PE; C57BL/6 mice; mLuc 0.75 mg/kg IV; n=3 | Readout: Approximately 3 h; excised-organ luminescence Comparator: Other screened ionizable lipidoids | 503O8,12 generated measurable reporter expression, with total organ signal on the order of 10^8 photons/s by graph inspection. This is an estimate, not an exact mean.Fig. 2b; six-organ sum |
| Spleen-biased reporter distribution Same 16PE formulation and IV dose; n=3 | Readout: Approximately 3 h; liver, spleen and lung signal fractions Comparator: Liver-biased library members | Spleen dominates the displayed three-organ signal distribution; previously reviewed graph estimate approximately 90%. This is a share of reporter luminescence, not percentage of injected RNA or transfected cells.Fig. 3a,c |
| Helper-lipid characterization 503O8,12 16PE, 40PE and 40TAP; n=4 | Readout: Particle size, encapsulation and zeta measurements Comparator: Three helper-lipid conditions | SI characterization supports a physical-property comparison. It does not establish 503O8,12 lung immune-cell selectivity or the toxicity ranking reported for 514O6,10.Previously reviewed SI Fig. S1 |
Mechanistic evidence
The paper links lipid chemistry and helper-lipid composition to expression distribution. Product-specific evidence for 503O8,12 supports spleen-biased reporter expression; a specific spleen cell target or receptor-mediated mechanism was not demonstrated.
5. Interpretation and unresolved details
- The in-depth lung NK/DC, endothelial and epithelial-cell experiments use 514O6,10, not 503O8,12.
- The paper’s 16PE versus 40TAP liver-enzyme and cytokine comparisons must not be represented as 503O8,12 safety data.
- Graph-derived estimates are approximate; do not present them as exact specifications.
- The three-organ expression fraction and six-organ total luminescence have different denominators.
- No product-specific therapeutic, CAR-T, repeat-dose or human efficacy claim is established.
- Full SI was reviewed in the earlier product record; fresh access to the SI file was restricted during this update. Exact numeric reanalysis awaits a downloadable source-data file.
Results apply to the stated LNP composition, cargo, process, dose, route and model. LNP experimental measurements are not batch-release specifications for the isolated lipid.
6. References and data access
Petersen DMS, Weiss RM, Hajj KA, et al. Branched-Tail Lipid Nanoparticles for Intravenous mRNA Delivery to Lung Immune, Endothelial, and Alveolar Cells in Mice. Advanced Healthcare Materials 13, e2400225 (2024).
Download experiment table (CSV) · Open printable Formulation & Performance guide
Revision FP-20261024-b2 · Source review: 24 October 2026 · DC Chemicals