Journal: Viruses

Article Title: Humoral Immune Response Profile of COVID-19 Reveals Severity and Variant-Specific Epitopes: Lessons from SARS-CoV-2 Peptide Microarray

doi: 10.3390/v15010248

Figure Lengend Snippet: Schematic representation of the study design, microarray protocol, and data analysis. ( a ) Sample acquisition and heat inactivation of virus, ( b ) SARS-CoV-2 whole proteome microarray design, ( c ) microarray staining and image acquisition, ( d ) data analysis pipeline.

Article Snippet: PEPperCHIP ® Peptide Microarray slides were brought to room temperature, assembled onto the PEPperCHIP ® incubation tray (PEPperPRINT GmbH, Germany), and equilibrated using the staining buffer for 15 min.

Techniques: Microarray, Virus, Staining

Journal: Viruses

Article Title: Humoral Immune Response Profile of COVID-19 Reveals Severity and Variant-Specific Epitopes: Lessons from SARS-CoV-2 Peptide Microarray

doi: 10.3390/v15010248

Figure Lengend Snippet: Heatmaps for IgA and IgG response showing major immunogenic regions identified in the SARS-CoV-2 whole proteome microarray. The printed proteome constitutes ORF1a/b polyprotein encoding 16 non-structural proteins (1–10 and 12–16), structural proteins (S, N, E, and M), and the accessory proteins (ORF3a, 6, 7a, 8, and 10).

Article Snippet: PEPperCHIP ® Peptide Microarray slides were brought to room temperature, assembled onto the PEPperCHIP ® incubation tray (PEPperPRINT GmbH, Germany), and equilibrated using the staining buffer for 15 min.

Techniques: Microarray

Journal: Bioengineering

Article Title: COVID-19 Diagnostic Strategies Part II: Protein-Based Technologies

doi: 10.3390/bioengineering8050054

Figure Lengend Snippet: The developed protein microarray-based tests for COVID-19 detection.

Article Snippet: PEPperPRINT GmbH [ ] , PEPperCHIP ® Pan-Corona Spike Protein Microarray , Antibodies against S antigen , S proteins derived from seven coronaviruses translated into overlapping peptides , (No info) , (No info) , One array with 4564 peptides in duplicate , RUO , For Serum antibody fingerprint analysis, Immune monitoring and Epitope studies.

Techniques: Microarray, Peptide Microarray, Bioprocessing, Derivative Assay, High Throughput Screening Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Virus, Clinical Proteomics

Journal: iScience

Article Title: Selection for immune evasion in SARS-CoV-2 revealed by high-resolution epitope mapping and sequence analysis

doi: 10.1016/j.isci.2023.107394

Figure Lengend Snippet: High-density peptide arrays (HDPA) provide high-resolution antibody epitope maps across the SARS-CoV-2 proteome (A) Overview of analytical pipeline. The proteome of SARS-CoV-2 was translated into 15-mer overlapping peptides with a peptide-to-peptide overlap of 13 amino acids. The resulting individual peptides were printed in duplicates on the microarray. Sera from confirmed SARS-CoV-2-positive and -negative individuals were incubated on PEPperCHIP the HDPA. Serum antibody binding was visualized using respective fluorescently labeled secondary antibodies (anti-human IgG and anti-human IgA). Image acquisition and data quantification resulted in epitope-specific antibody profiles for SARS-CoV-2. (B) Average relative fluorescent units (RFU) profiles and peptide coverages are plotted across the SARS-CoV-2 proteome (ORF1A, ORF1B, Spike (S) protein, Envelope (E) protein, Membrane (M) glycoprotein, Nucleocapsid (N) phosphoprotein). Antibody responses to each linear 15-mer peptide were mapped across the SARS-CoV-2 proteome and average RFU calculated for each amino acid residue. The normalized positional ‘epitope coverage’ at each protein residue location is defined as the ratio of total peptides mapped to each position by the total expected peptides (see section). ‘Hotspots’ can be seen as spiked in the RFU or coverage distributions. (C) Comparison of mean RFU (log-scale) between SARS-CoV-2-positive and -negative sample groups for each viral protein. (unpaired t-test, ns: p > 0.05; ∗: p ≤ 0.05; ∗∗: p ≤ 0.01; ∗∗∗: p ≤ 0.001; ∗∗∗∗: p ≤ 0.0001).

Article Snippet: To analyze the antibody responses to SARS-CoV-2 at the epitope level we used a recently developed high-density peptide array (HDPA), the PEPperCHIP Microarray (PEPperPRINT GmbH, Germany), covering the proteome of the SARS-CoV-2 isolate Wuhan-Hu-1 as well as the four seasonal hCoVs OC43, HKU1, NL63 and 229E (see for accession numbers used).

Techniques: Microarray, Incubation, Binding Assay, Labeling, Membrane, Residue, Comparison