pepperchip custom peptide microarray  (PEPperPRINT gmbh)

Journal: Frontiers in Immunology

Article Title: Mapping autoantibody targets of full-length C-reactive protein in systemic lupus erythematosus: importance for neutrophil function and classical complement activation

doi: 10.3389/fimmu.2025.1578372

Figure Lengend Snippet: Dot-plot between anti-CRP antibodies and epitope positivity. The graph displays anti-CRP antibodies versus the number of positive epitopes obtained with microarray-based linear epitope mapping in patients with systemic lupus erythematosus ( n =42). (CRP, C-reactive protein).

Article Snippet: The full sequence of the CRP monomer was printed in 15 a.a sequences with 14 a.a overlap using PEPperCHIP Custom Peptide Microarray (PEPperPRINT ® GmbH, Heidelberg, Germany).

Techniques: Microarray

Journal: Frontiers in Immunology

Article Title: Mapping autoantibody targets of full-length C-reactive protein in systemic lupus erythematosus: importance for neutrophil function and classical complement activation

doi: 10.3389/fimmu.2025.1578372

Figure Lengend Snippet: Heatmap representation of autoreactivity against CRP obtained with microarray-based linear epitope mapping of the full CRP monomer. (A, B) Individual signal intensity of IgG autoantibody reactivity against full-length CRP for subjects with SLE ( n =42) and HBD ( n =11). (C–E) Mean signal intensity of IgG autoantibody reactivity against motifs of full-length CRP comparing anti-CRP negative (anti-CRP–; n =16) vs. anti-CRP positive (anti-CRP+; n =26) patients with SLE, patients with ( n =6) and without ( n =36) active disease, and no damage ( n =20) vs. irreversible organ damage (any organ system; n =22). Each column represents one subject (A, B) or the mean value of a group (C–E) . Each row represents a 15 amino acid long sequence covering the full length of the protein with 14 amino acids overlap and 7 amino acid GS repeats elongated before and after the protein sequence. (CRP, C-reactive protein; HBD, healthy blood donors; SDI, Systemic Lupus International Collaborating Clinics/American College of Rheumatology damage index; SLE, systemic lupus erythematosus; SLEDAI-2K, SLE Disease Activity Index 2000).

Article Snippet: The full sequence of the CRP monomer was printed in 15 a.a sequences with 14 a.a overlap using PEPperCHIP Custom Peptide Microarray (PEPperPRINT ® GmbH, Heidelberg, Germany).

Techniques: Microarray, Sequencing, Activity Assay

Journal: Scientific Reports

Article Title: A high-throughput pipeline for design and selection of peptides targeting the SARS-Cov-2 Spike protein

doi: 10.1038/s41598-021-01225-2

Figure Lengend Snippet: Screening and selection of peptide binders to SARS-CoV-2 S protein. ( a ) Peptides P2-P11 represent wild-type versions of the original 27-mer ACE2 N-terminal alpha helix. They were designed as 18-mers spanning the length of the original fragment with 17 amino acids overlapped. The ACE2 fragment is predicted to bind to the SARS-CoV2 S protein via residues shown in bold . ( b ) Normalized binding signal of the ACE2-derived peptide variants show little to no binding to SARS-CoV2 S1 protein. There is an apparent trend for increased binding from the peptide fragments overlapping the center of the WT sequence and P7 shows the highest binding signal with a z-score of 1.3 when exposed to 50 µg/mL of S1 protein. ( c ) Normalized binding signal of the 14 peptides selected from microarray screening experiments for further characterization. 10 peptides were selected from the pool of ACE2 mutants, 3 were selected from the pool of modeled sequences, and one non-binding sequence was selected for comparison. All sequences selected had a Z-score > 2 on the 50 µg/mL S1 protein array, except for P481. ( d ) Sequences of the 14 peptides selected for synthesis with N-terminus biotin attached via a PEG4 spacer. **Note that P28 was not able to be synthesized by the vendor. ( e ) Binding curves of biotinylated peptides to immobilized SARS-CoV2 S1 protein in ELISA plate-based assay. Four peptides (P89, P100, P168 and P180) showed higher binding affinity than the original ACE2 fragment (SBP1) and were selected for further characterization.

Article Snippet: The library of 2,376 sequences was printed onto a custom PEPperCHIP peptide microarray (PEPperPrint GmbH, Heidelberg, Germany).

Techniques: Selection, Binding Assay, Derivative Assay, Sequencing, Microarray, Comparison, Protein Array, Synthesized, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: A high-throughput pipeline for design and selection of peptides targeting the SARS-Cov-2 Spike protein

doi: 10.1038/s41598-021-01225-2

Figure Lengend Snippet: Peptide Microarray to Identify SARS-CoV-2 S Protein Binding Sequences. ( a ) Schematic showing microarray screening procedure for detecting binding of a biotinylated target protein. ( b ) Microarray images of peptide subarray following exposure to 50, 10, or 5 µg/mL of SARS-CoV2 S1 protein. ( c ) Normalized binding signal of peptides after exposure to SARS-CoV2 S1 protein at 50, 10, or 5 µg/mL concentration. Z-score at each S1 concentration is plotted for peptides listed in numerical order along the X-axis. P2-P811 correspond to peptides derived from the ACE2 N-terminal alpha helix (approach 1) while P812-P2376 were derived from a random starting library and screened in silico for docking to S protein (approach 2). Peptides with high Z-scores (> 1.95, indicated by dotted line) represent S1 protein binding sequences.

Article Snippet: The library of 2,376 sequences was printed onto a custom PEPperCHIP peptide microarray (PEPperPrint GmbH, Heidelberg, Germany).

Techniques: Peptide Microarray, Protein Binding, Microarray, Binding Assay, Concentration Assay, Derivative Assay, In Silico