Table 3
Parameter: Aqueous phase | Reported procedure / limitation: mRNA in 25 mM citrate buffer, pH 4.4. | Source: Supporting Information p. 4
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iChol15-C4A2
Literature records
Search the tables and observations reproduced from the product literature guide. Rows may represent measurements, recipes or experimental context; they are not all independent experiments.
Table 3
Parameter: Organic phase | Reported procedure / limitation: The lipid mixture was dissolved in ethanol at 10 mg/mL. | Source: Supporting Information p. 4
Table 3
Parameter: Mixing | Reported procedure / limitation: Microfluidic mixing at aqueous:ethanol 3:1 by volume; total flow rate not reported. The formulation-optimization subsection also describes ethanol dilution. | Source: Supporting Information pp. 4–5
Table 3
Parameter: Lipid:mRNA loading | Reported procedure / limitation: iChol lipid:mRNA 25:1 by mass; this is the ionizable lipid mass, not the combined lipid mass and not N/P. | Source: Supporting Information pp. 4–5
Table 3
Parameter: Buffer exchange | Reported procedure / limitation: Overnight dialysis against PBS, pH 7.4, with a 20 kDa dialysis cassette. | Source: Supporting Information p. 4
Table 3
Parameter: Analytical methods | Reported procedure / limitation: DLS for hydrodynamic size and zeta potential; a modified RiboGreen assay for RNA encapsulation. | Source: Supporting Information p. 4
Table 3
Parameter: Unreported parameters | Reported procedure / limitation: No experimentally established N/P or target-specific apparent particle pKa was identified. | Source: Full article and Supporting Information review
Table 4
Measurement / batch: Initial formulation: size | Reported result: 90.76 ± 4.8 nm | Evidence context: Table S3, p. 60; 50:50:1.5 molar parts
Table 4
Measurement / batch: Initial formulation: PDI | Reported result: 0.088 | Evidence context: Table S3, p. 60; 50:50:1.5 molar parts
Table 4
Measurement / batch: Initial formulation: encapsulation | Reported result: 93.56 ± 1.06% | Evidence context: Table S3, p. 60; 50:50:1.5 molar parts
Table 4
Measurement / batch: Optimized Formulation 8: size | Reported result: 106 nm | Evidence context: Main Results accompanying Fig. 5c–e and SI Fig. S49; 60:50:1.5 molar parts
Table 4
Measurement / batch: Optimized Formulation 8: PDI / encapsulation | Reported result: 0.09 / 94.7% | Evidence context: Main Results accompanying Fig. 5c–e and SI Fig. S49
Table 5
Experiment and dose: Initial intramuscular reporter screen BALB/c mice; luciferase mRNA 5 µg/mouse; initial 50:50:1.5 formulation; n = 3 | Readout and comparator: Readout: 6 h Comparator: Experimental MC3 and ALC-0315 LNPs | Result and source: iChol15-C4A2 was among the leading local-expression candidates. This is injection-site expression, not an intravenous organ-targeting measurement. Main Fig. 5a–b; SI p. 4
Table 5
Experiment and dose: Intramuscular formulation optimization BALB/c mice; luciferase mRNA 1 µg/mouse; ten compositions; n = 3 | Readout and comparator: Readout: 6 h Comparator: Ten formulations spanning iChol 40–60, DSPC 30–60 and DMG-PEG2000 1.5–5 molar parts | Result and source: Formulation 8 (60:50:1.5) was selected for higher local reporter expression and carried forward. Main Fig. 5c–e; SI pp. 4–5 and Fig. S49
Table 5
Experiment and dose: Intravenous functional mRNA expression Mice; luciferase mRNA 0.1 mg/kg; optimized 60:50:1.5 formulation; n = 3 | Readout and comparator: Readout: 6 h Comparator: Experimental MC3 and ALC-0315 LNPs | Result and source: Spleen:liver luminescence ratio 4.78, versus 0.24 for MC3 and 0.12 for ALC-0315. The ratio is approximately 39.8 times the ALC-0315 ratio; this is not a 39.8-fold increase in absolute splenic expression. Main Fig. 5f–i and accompanying Results; SI p. 5
Table 5
Experiment and dose: Intravenous fluorescent RNA distribution BALB/c mice; Cy5-labelled mRNA 0.3 mg/kg; optimized formulation; n = 3 | Readout and comparator: Readout: 6 h Comparator: ALC-0315 LNPs | Result and source: The article describes an approximately 1.0 spleen:liver fluorescence ratio for Tc-LNPs versus approximately 0.2 for ALC-0315. Fluorescence tracks labelled material and does not establish translated protein or percentage of injected dose. Main Fig. 5j–l and accompanying Results; SI p. 5
Table 5
Experiment and dose: ApoE-dependent cellular uptake and expression HepG2 and HEK293 cells; LNPs preincubated with ApoE at 0–1 g ApoE/g total lipid for 15 min at 37 °C; 1 µg reporter mRNA | Readout and comparator: Readout: Cy5 uptake assessed after 2 h; expression assessed in the parallel reporter assay Comparator: ALC-0315 LNPs; with and without ApoE preincubation | Result and source: ApoE increased ALC-0315 uptake and expression in LDLR-expressing HepG2 cells, while the tested Tc-LNPs changed minimally. The comparison supports reduced dependence on ApoE-mediated uptake. Main Fig. 6a–f; SI pp. 5–6 and Figs. S51–S52
Table 5
Experiment and dose: Protein-corona analysis LNPs mixed 1:1 by volume with mouse plasma at 37 °C, then isolated and analysed by LC–MS | Readout and comparator: Readout: 1 h plasma incubation Comparator: ALC-0315 LNPs | Result and source: The tested Tc-LNP coronas contained less ApoE than the ALC-0315 comparator. Main-text and SI quantitative summaries are not identical, so no single exact ApoE percentage is assigned here. Main Fig. 6g–j; SI p. 6 and Tables S5–S7
Table 5
Experiment and dose: pH-dependent membrane disruption Mouse erythrocytes exposed to Tc-LNPs; physiological versus acidic buffer | Readout and comparator: Readout: 1 h at 37 °C Comparator: Physiological pH 7.4 and assay controls | Result and source: The tested Tc-LNPs showed greater membrane-disruptive activity under acidic conditions. The methods specify pH 5.5, whereas one Results passage uses pH 5.0; no exact acidic-pH percentage or pKa is assigned here. Main Fig. 7b; SI p. 6
Table 5
Experiment and dose: Storage of formulated RNA-loaded particles Optimized 60:50:1.5 luciferase-mRNA LNPs stored at 4 °C for 0, 1, 3, 7, 14 and 30 days; particle assays and cell reporter assays; IM test 5 µg/mouse, n = 3 | Readout and comparator: Readout: Up to 30 days; mouse reporter readout 6 h after injection Comparator: Experimental four-component MC3 and ALC-0315 LNPs | Result and source: The authors report retained physical properties, encapsulation and reporter activity over the tested storage period. This does not establish storage stability of neat lipid or empty LNPs. SI p. 7 and Fig. S57; main Results
Table 5
Experiment and dose: Short-term in vivo tolerability BALB/c mice; intravenous RNA-loaded Tc-LNPs, 0.5 mg mRNA/kg | Readout and comparator: Readout: 24 h Comparator: PBS control | Result and source: No significant changes in ALT, AST, BUN or creatinine, or obvious major-organ histopathology, were reported under the tested conditions. This is a short-term mouse observation. Main Fig. 7d–e; SI p. 7