human galectin 3 binding protein  (Galectin Therapeutics)

Journal: Nature Communications

Article Title: SARS-CoV-2 RNAemia and proteomic trajectories inform prognostication in COVID-19 patients admitted to intensive care

doi: 10.1038/s41467-021-23494-1

Figure Lengend Snippet: a Magnetic bead-based affinity isolation of binding partners using His-tagged SARS-CoV-2 spike glycoprotein as a bait for proteins in SARS-CoV-2-positive patient plasma ( n = 8). Volcano plot depicting significantly enriched constant chains of immunoglobulins. b Volcano plot depicting significantly enriched non-immunoglobulin proteins ( n = 8). c Comparison of SARS-CoV-2 spike glycoprotein pulldown using plasma from COVID-19 ICU patients ( n = 8) and non-COVID-19 patients ( n = 3). Significance was determined by paired Student’s t tests for ( a ) and ( b ) and unpaired Student’s t tests for ( c ). d LGALS3BP levels across three patient cohorts as determined by DIA-MS or ELISA: control patients before undergoing elective cardiac surgery ( n = 30), pre-pandemic sepsis ICU patients ( n = 12) and COVID-19 ICU patients ( n = 74). Kruskal–Wallis and Dunn’s multiple comparisons tests were used to determine statistical significance. e Volcano plot representing protein changes between baseline plasma samples from patients in ICU with either sepsis ( n = 12) or COVID-19 ( n = 12). Significance was determined through the Mann–Whitney U test with Benjamini-Hochberg’s FDR correction. f Plasma proteins correlating to LGALS3BP after age and sex corrections in COVID-19 ICU patients ( n = 12) are highlighted by a Spearman correlation matrix across the proteomic dataset. Proteins with a Spearman correlation coefficient greater than 0.5 were used for gene ontology pathway enrichment analysis (Supplementary Fig. ). All statistical analyses are two-tailed.

Article Snippet: ELISAs for receptor for advanced glycation end-products (RAGE; R&D Systems, DRG00), galectin-3-binding protein (LGALS3BP; R&D Systems, DGBP30B), and pentraxin-3 (PTX3; Abcam, ab214570) were performed according to the manufacturer’s instructions.

Techniques: Isolation, Binding Assay, Comparison, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Two Tailed Test

Journal: Nature Communications

Article Title: SARS-CoV-2 RNAemia and proteomic trajectories inform prognostication in COVID-19 patients admitted to intensive care

doi: 10.1038/s41467-021-23494-1

Figure Lengend Snippet: a Schematic representation of the SARS-CoV-2 spike-mediated cell-cell fusion assay. b–d Vero and HEK293-ACE2 cells were transfected either with pcDNA3 (plasmid backbone), pmCherry (plasmid coding mCherry), pLGALS3BP (plasmid coding LGALS3BP), siACE2 (siRNA targeting ACE2), or siNT1 (non-targeting siRNA), followed by transfection of pAAV-Spike (plasmid coding SARS-CoV-2 spike) 24 h later. After 20 h, cells were stained with anti-LGALS3BP (violet), anti-Spike (green), and DAPI for nuclei (blue). Representative images are shown in ( b ), and quantifications are shown in ( c) for Vero cells and in ( d ) for HEK293-ACE2 cells. Data (mean ± standard deviation; n = 6; Mann–Whitney U test) are plotted as the percentage of fused cells (syncytia) normalized to the total number of cells. Scale bars in ( b ) represent 100 µm. e Schematic representation of the SARS-CoV-2 spike/VSV-G pseudoparticle transduction assay. f – h HEK293-ACE2 cells were transfected either with pcDNA3, pLGALS3BP, siACE2, or siNT1, followed by the addition of spike- or VSV-G pseudoparticles carrying a GFP reporter 24 h later. After 36 h, cells were stained with anti-LGALS3BP (red), anti-GFP (green), and DAPI for nuclei (blue). Representative images are shown in ( f ) and quantifications are shown in ( g ) for spike-pseudoparticles (mean ± standard deviation; n = 6; Mann–Whitney U test) and in ( h ) for VSV-G pseudoparticles (mean ± standard deviation; n = 3). Data are plotted as the percentage of GFP-positive cells normalized to the total number of cells. Scale bars in ( f ) represent 200 µm. All statistical analyses are two-tailed.

Article Snippet: ELISAs for receptor for advanced glycation end-products (RAGE; R&D Systems, DRG00), galectin-3-binding protein (LGALS3BP; R&D Systems, DGBP30B), and pentraxin-3 (PTX3; Abcam, ab214570) were performed according to the manufacturer’s instructions.

Techniques: Cell-Cell Fusion Assay, Transfection, Plasmid Preparation, Staining, Standard Deviation, MANN-WHITNEY, Transduction, Two Tailed Test