Journal: Journal of the American Society for Mass Spectrometry

Article Title: Tracking the Behavior of Monoclonal Antibody Product Quality Attributes Using a Multi-Attribute Method Workflow.

doi: 10.1021/jasms.0c00432

Figure Lengend Snippet: Figure 5. NPD workflow for HCP and sequence variant analysis. (a) HCP analysis was carried out by spiking cathepsin L and rhLPL at different concentrations into a commercially available IgG1. (b) Sequence variant analysis was carried out using the same commercially available IgG1 originator and an in-house produced biosimilar. For both analyses, the samples were digested with trypsin, and analysis was carried out as previously described.

Article Snippet: Recombinant mouse cathepsin L protein and recombinant human lipoprotein lipase (rhLPL) protein and CF were purchased from R&D Systems (Abingdon, United Kingdom) and used for host cell protein (HCP) spiking experiments.

Techniques: Sequencing, Variant Assay, Produced

Journal: MedComm

Article Title: Outer membrane vesicle contributes to the Pseudomonas aeruginosa resistance to antimicrobial peptides in the acidic airway of bronchiectasis patients

doi: 10.1002/mco2.70084

Figure Lengend Snippet: Lactate‐mediated acidification dampens the bactericidal activity of LL‐37 against P. aeruginosa . (A) Lactate concentrations of the sputa were collected from bronchiectasis (BE) patients and their relatives as controls in cohort 3. Induced sputa were collected when needed. The comparison was performed with an unpaired Student t ‐test. (B) Comparison of the sputum LL‐37 concentrations between BE patients with different microbiology test results in cohort 3. A comparison was performed with one‐way analysis of variance (ANOVA) with Welch's test. (C) The lactate concentrations in bronchoalveolar lavage fluid (BALF) from mice with or without P. aeruginosa chronic lung infection. A comparison was performed with an unpaired Student t ‐test. (D) The survivals of three clinical P. aeruginosa isolated from BE patients after the treatment with LL‐37 in vitro. Comparison between the untreated and LL‐37‐treated groups (120 µg/mL) was performed with an unpaired Student t ‐test. (E) Percentages of survived clinical P. aeruginosa isolate SHPHC2 post‐LL‐37 treatment in the conditions with various pH adjusted by lactate. One‐way ANOVA with Welch's test was used for statistical analysis in both LL37‐treated and untreated groups, p ‐values were indicated in the figure, and results with a p ‐value less than 0.05 were considered statistically significant. The error bars plotted indicate the standard error of the mean.

Article Snippet: The final concentrations of LL‐37 (Tocris Bioscience, cat. 5213), P. aeruginosa ‐derived OMVs, HHQ (TargetMol, cat. T19713), PQS (TargetMol, cat. T38373) and Cl‐adimine (MedChemExpress, cat. HY‐100574) were stated in the main text.

Techniques: Activity Assay, Comparison, Infection, Isolation, In Vitro

Journal: MedComm

Article Title: Outer membrane vesicle contributes to the Pseudomonas aeruginosa resistance to antimicrobial peptides in the acidic airway of bronchiectasis patients

doi: 10.1002/mco2.70084

Figure Lengend Snippet: P. aeruginosa produces outer membrane vesicles (OMVs) to resist LL‐37‐mediated killing. (A) Quantification of OMVs produced by P. aeruginosa in the lactate‐mediated acidic environment by measuring FM 4‐64 fluorescence. One‐way analysis of variance (ANOVA) with Welch's test was used for statistical analysis. (B) Nanoparticle tracking analysis and transmission electron microscopy of OMVs isolated from P. aeruginosa overnight culture by ultracentrifugation. (C) Survival of P. aeruginosa PAO1 post‐LL‐37 treatment with the additions of OMVs of different dosages. The OMVs added were quantified by measuring the total proteins enwrapped. One‐way ANOVA with Welch's test was used for statistical analysis in both LL37‐treated and untreated groups and the p ‐value was shown. (D) The role of OMVs in the acidic condition‐induced strengthening of P. aeruginosa LL‐37 resistance was determined by adding Cl‐adimine to the bacteria‐killing system. The final concentration of Cl‐diamine was indicated on the diagram. The differences between groups were statistically analyzed by using one‐way ANOVA with Welch's test, with the p values of multiple comparisons indicated in the figures. Results with a p ‐value less than 0.05 were considered statistically significant. The error bars plotted indicate the standard error of the mean.

Article Snippet: The final concentrations of LL‐37 (Tocris Bioscience, cat. 5213), P. aeruginosa ‐derived OMVs, HHQ (TargetMol, cat. T19713), PQS (TargetMol, cat. T38373) and Cl‐adimine (MedChemExpress, cat. HY‐100574) were stated in the main text.

Techniques: Membrane, Produced, Fluorescence, Transmission Assay, Electron Microscopy, Isolation, Bacteria, Concentration Assay

Journal: Expert opinion on drug delivery

Article Title: Treprostinil palmitil inhalation powder leverages endogenous lung enzymes to provide sustained treprostinil.

doi: 10.1080/17425247.2024.2395444

Figure Lengend Snippet: Figure 4. Conversion of TP to TRE and expression of LPL by human lung cells. (a) The conversion of TP to TRE by various types of human lung cells. Cultured human lung cells were treated with 10 µm TP for 24 h. The TRE detected in each culture medium sample was corrected for non-enzymatic TP conversion (in the absence of cells) and normalized to the cell count of the same sample. Cell count per sample (million cells) was 1.3 ± 0.1 for A-549, 1.1 ± 0.2 for HASMC, 0.2 ± 0.2 for HLF, 0.9 ± 0.2 for HPAEC, 1.6 ± 0.2 for HSAEC, and 0.6 ± 0.4 for THP-1. N = 3 samples per group. (b) Baseline LPL expression (~53.1 kDa) by human lung cells determined by Western Blot. Each sample was run in triplicate. A-549: alveolar epithelium-like A-549 cells; HASMC: human artery smooth muscle cells; HLF: human lung fibroblasts; HPAEC: human pulmonary artery endothelial cells; HSAEC: human small airway epithelial cells; THP-1: macrophage-like THP-1 cells; hLPL: recombinant human LPL expressed in Chinese hamster ovary cells.

Article Snippet: Recombinant human LPL expressed in CHO cells (9888-LL-100, R&D Systems) was used at 10 μg/mL as the positive control.

Techniques: Expressing, Cell Culture, Cell Counting, Western Blot, Recombinant