Journal: Nature Communications

Article Title: Poly(GR) and poly(GA) in cerebrospinal fluid as potential biomarkers for C9ORF72 -ALS/FTD

doi: 10.1038/s41467-022-30387-4

Figure Lengend Snippet: a In spike-and-recovery experiments, recombinant (GR) 8 peptide was added to CSF from healthy individuals and subsequently serially diluted into a separate tube containing CSF from healthy individuals to create a 5-point series. The average recovery rate in two different samples was 81–88%. b The assay can detect purified (GR) 8 peptide at concentrations as low as 3 pg/ml. Replicates of 8-point series standard curve run on different dates are shown. A four-parameter logistic curve was used to fit the dose-response with GraphPad Prism 9.1. c Across the dynamic range of the assay, the coefficient of variation is less than 16% for all points except one concentration (100 pg/ml), below the 20% cutoff recommended by the FDA in standard variability for liquid-binding assays. d Dilution linearity was evaluated by measuring duplicates of 5 dilutions of 2 patient CSF samples spiked with 200 ng/ml (GR) 8 peptide. e Poly(GR) immunoassay specifically detects (GR) 8 peptide in control CSF and did not cross react with even up to 100 ng of (GP) 8 peptide in control CSF, n = 3 independent experiments, values are mean ± s.e.m. f CSF samples from 4 healthy and 4 patients with C9ORF72 mutations from the Mayo Clinic, Florida, values are mean ± s.e.m ( p = 0.0017, Unpaired t -test with Welch’s correction). g CSF samples from 3 healthy people from the NEALS cohort and 5 CSF samples from two patients with C9ORF72 mutations from the Barrow Institute, Arizona, values are mean ± s.e.m ( p = 0.0012, Unpaired t -test with Welch’s correction). h 19 healthy people, 19 patients with ALS but without C9ORF72 mutations (3 patients with duplicate samples, 16 patients with single samples), and 10 patients with C9ORF72 mutations (8 patients with duplicate samples, 2 patients with single samples) from the NEALS Consortium. Values from duplicate samples are presented as mean ± s.e.m ( p = 0.9460 control vs non-C9-ALS, p < 0.0001 non-C9-ALS vs C9-ALS, p < 0.0001 control vs C9-ALS Ordinary one-way ANOVA-Tukey’s multiple comparisons test). Source data are provided as a Source Data file.

Article Snippet: A four-parameter logistic curve was used to fit the dose-response with GraphPad Prism 9.1. c Across the dynamic range of the assay, the coefficient of variation is less than 16% for all points except one concentration (100 pg/ml), below the 20% cutoff recommended by the FDA in standard variability for liquid-binding assays. d Dilution linearity was evaluated by measuring duplicates of 5 dilutions of 2 patient CSF samples spiked with 200 ng/ml (GR) 8 peptide. e Poly(GR) immunoassay specifically detects (GR) 8 peptide in control CSF and did not cross react with even up to 100 ng of (GP) 8 peptide in control CSF, n = 3 independent experiments, values are mean ± s.e.m. f CSF samples from 4 healthy and 4 patients with C9ORF72 mutations from the Mayo Clinic, Florida, values are mean ± s.e.m ( p = 0.0017, Unpaired t -test with Welch’s correction). g CSF samples from 3 healthy people from the NEALS cohort and 5 CSF samples from two patients with C9ORF72 mutations from the Barrow Institute, Arizona, values are mean ± s.e.m ( p = 0.0012, Unpaired t -test with Welch’s correction). h 19 healthy people, 19 patients with ALS but without C9ORF72 mutations (3 patients with duplicate samples, 16 patients with single samples), and 10 patients with C9ORF72 mutations (8 patients with duplicate samples, 2 patients with single samples) from the NEALS Consortium.

Techniques: Recombinant, Purification, Concentration Assay, Binding Assay

Journal: Medical Microbiology and Immunology

Article Title: Quantitative assay to analyze neutralization and inhibition of authentic Middle East respiratory syndrome coronavirus

doi: 10.1007/s00430-024-00789-w

Figure Lengend Snippet: Comparison of MERS-CoV MNA to VNT 100 . a Exemplary inhibition curves of the antibody m336, a negative human serum, and the MERS-CoV WHO international standard for immunoglobulins (WHO IS, human serum) in the microneutralization assay (MNA). The logarithmic concentration of m336 or the logarithmic serum dilution is plotted against the foci reduction in % of the virus only control (VOC). R 2 values from the 4PL regression analysis and the calculated IC 50 values are noted on the graph. The dotted line visualizes the 50% inhibition compared to VOC. Repeated analysis of m336 ( b ) and WHO IS ( c ) in the MNA and the VNT 100 to calculate IC 50 and IC 80 values (MNA) or IC 100 values (VNT 100 ). d Correlation of the IC 50 values generated with the MNA and the IC 100 values of the VNT 100 . The line reflects the best-fit linear relationship between the variables, and the dotted lines represent 95% confidence intervals. p and r values reflect two-tailed Spearman’s rank-correlation tests. The LLODs of each test are visualized by the gray lines

Article Snippet: These values were used to interpolate 50% or 80% serum neutralization titer (NT 50 or NT 80 ) or inhibitory concentration (IC 50 or IC 80 ) by four-parameter logistic (4PL) regression analysis (GraphPad Prism 9 Software, La Jolla, California).

Techniques: Comparison, Inhibition, Microneutralization Assay, Concentration Assay, Virus, Control, Generated, Two Tailed Test

Journal: Medical Microbiology and Immunology

Article Title: Quantitative assay to analyze neutralization and inhibition of authentic Middle East respiratory syndrome coronavirus

doi: 10.1007/s00430-024-00789-w

Figure Lengend Snippet: Inhibition of MERS-CoV by the pan-CoV inhibitor EK1C4. The inhibition curves of the antibody m336 ( a ) and EK1C4 ( b ) in the MNA. The logarithmic concentration of m336 or the molarity of EK1C4 is plotted against the foci reduction in % of the virus only control (VOC). R 2 values from the 4PL regression analysis and the calculated IC 50 values are noted on the graph

Article Snippet: These values were used to interpolate 50% or 80% serum neutralization titer (NT 50 or NT 80 ) or inhibitory concentration (IC 50 or IC 80 ) by four-parameter logistic (4PL) regression analysis (GraphPad Prism 9 Software, La Jolla, California).

Techniques: Inhibition, Concentration Assay, Virus, Control