Journal: Cell Reports Medicine

Article Title: PARP-1 improves leukemia outcomes by inducing parthanatos during chemotherapy

doi: 10.1016/j.xcrm.2023.101191

Figure Lengend Snippet:

Article Snippet: Human: HS-5 stromal cells , LGC Standards , ATCC Cat# CRL-11882, RRID: CVCL_3720.

Techniques: Recombinant, Derivative Assay, Staining, Negative Control, Plasmid Preparation, Caspase-3 Assay, TUNEL Assay, Reverse Transcription, Expressing, Bicinchoninic Acid Protein Assay, Western Blot, Microarray, Software, Fluorescence

Journal: Electrophoresis

Article Title: Development of an imaged capillary isoelectric focusing method for characterizing the surface charge of mRNA lipid nanoparticle vaccines

doi: 10.1002/elps.201900063

Figure Lengend Snippet: (A) Electropherograms of an LNP using various ampholytes and additives. Traces A and B show high background or precipitation and LNP containing sharp peaks using the broad range Servalyt pH 2–9 and Pharmalyte ampholytes pH 3–10, respectively. Trace C shows a focused LNP with an apparent pI of approximately 7.3 using pH 3–10 Pharmalyte ampholytes containing 10% glycerol. Trace D uses a mixture of 33.3% ampholyte pH 5–8 and 66.6% ampholytes pH 3–10 with 10% glycerol. Trace E uses a 66.6% ampholyte pH 5–8 and 33.3% ampholytes pH 3–10 with 10% glycerol. Trace F uses ampholyte pH 5–8 containing 10% glycerol. The pI of the LNP shifts to approximately 7.6–7.8 in traces D, E, and F. Two pI markers are 5.85 and 8.40. (B) Electropherogram of an LNP prepared in triplicate. An LNP sample was prepared in triplicate for the icIEF experiment. The LNP has an apparent pI of approximately 7.89 and peak shape was consistent for the three replicates. (C) Calibration curve of LNP, which ranges from 7.2–115 µg/mL of total lipids. LNP samples were diluted in icIEF ampholyte mixtures from 0.56 to 9.0 µg/mL of mRNA (equivalent to 7.2 to 115 µg/mL of total lipid). This linear range has a coefficient of determination ( R 2 ) ≥ 0.997.

Article Snippet: All methylcellulose containing solutions, Maurice icIEF fluorescence calibration standards, system suitability standards, pI markers (5.85 and 8.40), Servalyt pH 2–9 ampholytes, and icIEF cartridges were obtained from ProteinSimple (Santa Clara, CA, USA).

Techniques:

Journal: Electrophoresis

Article Title: Development of an imaged capillary isoelectric focusing method for characterizing the surface charge of mRNA lipid nanoparticle vaccines

doi: 10.1002/elps.201900063

Figure Lengend Snippet: LNP pI plotted by cationic lipids and mRNA concentration: different LNP batches were formulated to contain different cationic lipid to mRNA ratios. These LNPs were then diluted to five different lipid concentrations and subjected to icIEF. The pI ( y ‐axis) was plotted against cationic lipid concentration (Fig. A) and the mRNA concentration (Fig. B). A strong correlation of pI to cationic lipid concentration was observed ( R = 0.956; logarithmic fit, left graph) compared to a weaker correlation of pI to mRNA concentration ( R = 0.653; logarithmic fit, right graph).

Article Snippet: All methylcellulose containing solutions, Maurice icIEF fluorescence calibration standards, system suitability standards, pI markers (5.85 and 8.40), Servalyt pH 2–9 ampholytes, and icIEF cartridges were obtained from ProteinSimple (Santa Clara, CA, USA).

Techniques: Concentration Assay

Journal: Electrophoresis

Article Title: Development of an imaged capillary isoelectric focusing method for characterizing the surface charge of mRNA lipid nanoparticle vaccines

doi: 10.1002/elps.201900063

Figure Lengend Snippet: (A) UV absorbance of LNPs in both aqueous and icIEF ampholyte mixtures. LNPs containing mRNA were tested in Tris buffer (blue trace) and icIEF ampholyte mixture (black dash trace). LNPs without mRNA were tested in Tris buffer (green trace) and icIEF ampholyte mixture (red dash trace). LNPs containing mRNA show an absorbance max at 260 nm compared to LNP without mRNA, which lack a peak at 260 nm. Both aqueous and icIEF ampholyte mixtures absorbance traces were identical when comparing the wavelengths at 260 nm demonstrating that with or without mRNA, LNPs are stable and intact in the final cIEF ampholyte mixture. (B) Electropherogram of LNP formulated with and without mRNA. LNPs without mRNA (red dashed trace) have a similar pI to LNPs formulated with mRNA (black solid trace). The pI of both LNPs is approximately 7.6–7.7. Two pI markers were 5.85 and 8.40.

Article Snippet: All methylcellulose containing solutions, Maurice icIEF fluorescence calibration standards, system suitability standards, pI markers (5.85 and 8.40), Servalyt pH 2–9 ampholytes, and icIEF cartridges were obtained from ProteinSimple (Santa Clara, CA, USA).

Techniques: