protoarraytm human protein microarrays v5.1  (Thermo Fisher)

Journal: The Journal of Clinical Investigation

Article Title: Prevalence and pathogenicity of autoantibodies in patients with idiopathic CD4 lymphopenia

doi: 10.1172/JCI136254

Figure Lengend Snippet: Sera from 51 ICL patients and 25 HCs were screened for autoantibodies using a high-throughput 124 autoantigen microarray platform. (A) Volcano plots of the IgG and IgM autoantibodies, on the left and the right, respectively, displaying –log10(P value) on the y axis versus log2 (average Ab score in ICL samples/average Ab score in HC samples) on the x axis. Each circle represents an autoantibody, highlighting in blue (IgG) or green (IgM) the statistically significant positive autoantibodies between the HC and ICL groups, calculated with the nonparametric Mann-Whitney test with Bonferroni’s correction. Only targets having P < 0.05/122 (with 122 being the number of comparisons) were considered significant and are highlighted. (B) Venn diagram showing antigens recognized by both IgG and IgM (pink) vs. by only IgG (blue) or only IgM (green) autoantibodies. (C) Number of autoantibody targets with Z ≥ 4 for HCs and each subgroup of ICL patients. Z scores for each target were calculated as the number of standard deviations the Ab score was above the mean of the HC Ab score for of each target. Group 1 (open circles, n = 22) corresponds to ICL patients without a diagnosed autoimmune disease and without a positive test for a set of clinical autoantibodies. Group 2 (cyan circles, n = 15) corresponds to patients who tested positive for clinical autoantibodies but did not meet clinical criteria for any specific autoimmune diagnosis. Group 3 (blue circles, n = 14) corresponds to patients who had been diagnosed with 1 or more autoimmune disease. Data were pooled from 2 independent experiments. **P < 0.01; ***P < 0.001; ****P < 0.0001 by Kruskal-Wallis test and Dunn’s correction for multiple comparisons.

Article Snippet: The sera were then incubated on the Human Protein Microarray v5.1 (Thermo Fisher Scientific) displaying more than 9,000 human proteins.

Techniques: High Throughput Screening Assay, Microarray, MANN-WHITNEY

Journal: The Journal of Clinical Investigation

Article Title: Prevalence and pathogenicity of autoantibodies in patients with idiopathic CD4 lymphopenia

doi: 10.1172/JCI136254

Figure Lengend Snippet: Sera from ICL patients (n = 34) and HCs (n = 15) were screened for the presence of autoantibodies using the Human Protein Microarray v5.2. Sera were incubated on a microarray that displays over 9,000 full-length purified human proteins in their native conformations. Data were batch corrected, filtered for relative fluorescence units (RFUs) >500 for at least 1 sample for each particular protein, and normalized. The Z score for each target was calculated as the number of standard deviations the signal for a specific target had above the mean of the HCs. (A) Proportion of HC (gray) or ICL patients (blue) that had IgG Abs at Z scores ≥3, ≥4, or ≥5. (B) Number of proteins targeted by IgG Abs with Z score ≥4 from individual HC (gray) or patients’ sera grouped according to autoimmune status, as described in Figure 1C. (C) Percentage of participants (HC, gray; ICL, blue) that shared any of the 2,159 targets found at Z ≥ 4. Data were pooled from 3 independent experiments. **P < 0.01 by Kruskal-Wallis test and Dunn’s correction for multiple comparisons.

Article Snippet: The sera were then incubated on the Human Protein Microarray v5.1 (Thermo Fisher Scientific) displaying more than 9,000 human proteins.

Techniques: Microarray, Incubation, Purification, Fluorescence