Journal: Frontiers in Immunology

Article Title: Complement activation and cellular inflammation in Fabry disease patients despite enzyme replacement therapy

doi: 10.3389/fimmu.2024.1307558

Figure Lengend Snippet: Serum concentrations of C3a and C5a in FD patients. (A) C3a and (B) C5a concentrations (ng/ml) in serum samples from healthy controls (n=18) and treatment-naiüve (n=24) FD patients. (C) C3a and (D) C5a concentrations (ng/ml) in serum samples from healthy controls (n=18), treatment-naiüve FD patients and from the same patients under ERT (n=17). Data were tested for normal distribution by Kolmogorov-Smirnov test. Differences between groups were determined by Mann-Whitney test (A, B) when data were not normally distributed or Ordinary one-way ANOVA with Tukey’s posthoc test (C, D) in case of normal distribution. Data in A, B are shown as median and range with 25th and 75th percentile. Data in (C, D) are shown as mean + SEM (C, D) . *p<0.05, **p<0.01; ***p<0.001; ****p<0.0001. (E, F) Percentage of serum samples from healthy controls (white bar, n=18), treatment-naiüve FD patients (black bar, n=17) and from the same patients under ERT (grey bar, n=17) with levels of (E) ≤5,000 ng/ml or >5,000 ng/ml of C3a or (F) ≤20 ng/ml, 21-35 ng/ml or >35 ng/ml C5a in serum. (G, H) Linear regression analysis of C5a and C3a serum levels (ng/ml) in FD patients before (G) and after (H) ERT (n=17). R 2 is the coefficient of determination of linear regression and ranges from 0 (no linear regression) to 1 (perfect linear regression).

Article Snippet: For quantification of complement cleavage products C3a/C3a-desArg (C3a) as well as C5a/C5a-desArg (C5a) in serum, commercial C3a (HK354, Hycult Biotech, Uden, Netherlands) and C5a (HK349, Hycult Biotech, Uden, Netherlands) human enzyme-linked immunosorbent assay (ELISA) kits were used according to the manufacturer’s instructions.

Techniques: MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Complement activation and cellular inflammation in Fabry disease patients despite enzyme replacement therapy

doi: 10.3389/fimmu.2024.1307558

Figure Lengend Snippet: C3a, C5a and lyso-Gb3 concentrations in the circulation of FD patients with missense/nonsense mutations. (A–D) C3a, (E–H) C5a and (I–L) lyso-Gb3 concentrations (ng/ml) in samples from (A, E, I) treatment-naiüve (n=17), (B, F, J) FD patients under ERT (n=17) with missense (n=9) or nonsense (n=8) GLA mutations and from FD patients with (C, G, K) missense (n=9) or (D, H, L) nonsense (n=8) GLA mutations before (V0) or under ERT (V1). Data were tested for normal distribution by Kolmogorov-Smirnov test. Differences between groups were determined by unpaired t-test (A, B, E, F, I, J ) or paired t- test (C, D, G, H, K, L) . Data in (A, B, E, F, I, J) are shown as mean + SEM. *p<0.05; ***p<0.001.

Article Snippet: For quantification of complement cleavage products C3a/C3a-desArg (C3a) as well as C5a/C5a-desArg (C5a) in serum, commercial C3a (HK354, Hycult Biotech, Uden, Netherlands) and C5a (HK349, Hycult Biotech, Uden, Netherlands) human enzyme-linked immunosorbent assay (ELISA) kits were used according to the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Immunology

Article Title: Complement activation and cellular inflammation in Fabry disease patients despite enzyme replacement therapy

doi: 10.3389/fimmu.2024.1307558

Figure Lengend Snippet: C3a, C5a and lyso-Gb3 concentrations in the circulation of FD patients with ADA-negative/positive status. (A–D) C3a, (E–H) C5a and (I–L) lyso-Gb3 concentrations (ng/ml) in serum from ADA-negative (n=6) or ADA-positive (n=11) (A, E, I) patients, who were either treatment-naiüve (n=17) or (B, F, J) received ERT (n=17). (C, G, K) ADA-negative (n=6) or (D, H, L) ADA-positive (n=11) FD patients before (V0) or under ERT (V1). Red lines denote patients with missense mutations. Data were tested for normal distribution by Kolmogorov-Smirnov test. Differences between groups were determined by unpaired t-test (A, B, E, F, I, J) or paired t-test (C, D, G, H, K, L) . Data in (A, B, E, F, I, J) are shown as mean + SEM. *p<0.05; ***p<0.001.

Article Snippet: For quantification of complement cleavage products C3a/C3a-desArg (C3a) as well as C5a/C5a-desArg (C5a) in serum, commercial C3a (HK354, Hycult Biotech, Uden, Netherlands) and C5a (HK349, Hycult Biotech, Uden, Netherlands) human enzyme-linked immunosorbent assay (ELISA) kits were used according to the manufacturer’s instructions.

Techniques:

Journal: iScience

Article Title: sMR and PTX3 levels associate with COVID-19 outcome and survival but not with Long COVID

doi: 10.1016/j.isci.2024.110162

Figure Lengend Snippet:

Article Snippet: C3a, Human ELISA Kit , Hycult Biotech , Cat# HK354.

Techniques: Enzyme-linked Immunosorbent Assay, Software, Luminex

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Large-Scale Plasma Proteome Epitome Profiling is an Efficient Tool for the Discovery of Cancer Biomarkers

doi: 10.1016/j.mcpro.2023.100580

Figure Lengend Snippet: Mimotope heterogeneity and epitopes of human complement system components. SPR experiment ( A ) with BSI0442 mAb (factor H) and one of it’s cognate, biotinylated mimotope peptide (#5, immobilized). Binding inhibition curves obtained by competition with pooled human plasma ( left ) or natural CFH purified from plasma ( right ). Domain structure ( B ) of the CFH, CFHL1 and CFHR proteins, the N-terminal domains are shown in yellow . (C) The CFH concentration (µg/ml) of 37 samples was determined using selected BSI mAbs and a commercial CFH kit (Hycult) in a sandwich ELISA setup. Data for three pairwise comparisons in which seven individual samples were randomly chosen and colored (three red , four green ). Binding test summary ( D ) of QP mAbs that recognize CFH. mAb binding was tested on complete (FH), N-terminal (FH1-4), and C-terminal (FH15–20) fragments of factor H protein and on factor H related proteins (CFHR-1, CFHR-4B, and CFHR-5).

Article Snippet: The CFH concentration of selected samples was measured using a commercially available complement factor H, human, ELISA kit (Hycult Biotech), following the manufacturer’s protocol.

Techniques: Binding Assay, Inhibition, Purification, Concentration Assay, Sandwich ELISA