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Pepscan Inc spike protein peptide microarray
a Antibody-mediated neutralization of infection of luciferase-encoding VSV particles pseudotyped with spike proteins of MERS-CoV, SARS-CoV, SARS-CoV-2 and HCoV-OC43. Pseudotyped VSV particles pre-incubated with antibodies at indicated concentrations were used to infect VeroCCL81 cells (MERS-S pseudotyped VSV), VeroE6 cells (SARS-S and SARS2-S pseudotyped VSV) or HRT-18 cells (OC43-S pseudotyped VSV) and luciferase activities in cell lysates were determined at 20 h post transduction to calculate infection (%) relative to non-antibody-treated controls. The average ± SD ( n ≥ 6) from at least two independent experiments performed is shown. Iso-CTRL: an anti-Strep-tag human monoclonal antibody was used as an antibody isotype control. The IC 50 and IC 90 values were shown. Source data are provided as a Source Data file. b Antibody-mediated neutralization of MERS-CoV, SARS-CoV and SARS-CoV-2 infection. Neutralization of authentic viruses was performed using a plaque reduction neutralization test (PRNT) on VeroCCL81 cells (MERS-CoV) or VeroE6 (SARS-CoV and SARS-CoV-2) as described earlier  ,  . The experiment was performed with triplicate samples, the average ± SD is shown. The IC 50 and IC 90 values were shown. Source data are provided as a Source Data file. c ELISA-based receptor-binding inhibition assay. MERS-S ecto pre-incubated with serially diluted mAbs was added to ELISA plates coated with soluble human DPP4. The binding of MERS-S ecto to DPP4 was detected using an HRP-conjugated antibody recognizing the C-terminal Strep-tag on MERS-S ecto . Data points represent the average ± SD, for n = 3 replicates from two independent experiments. Source data are provided as a Source Data file. d Cell-cell fusion inhibition assay. Huh-7 cells—transfected with plasmid expressing (GFP-tagged) MERS-CoV S were pre-incubated in the presence or absence of 1.6C7 and 28D9, or an irrelevant iso-type control antibody (Iso-CTRL), and then treated with trypsin to activate the membrane fusion function of the MERS-CoV S protein. The formation of MERS-S mediated syncytia was visualized by fluorescence microscopy. Merged images of MERS-S-GFP expressing cells (green) and DAPI-stained cell nuclei (blue) are shown. The experiment was performed twice, data from a representative experiment is shown. Source data are provided as a Source Data file.
Spike Protein Peptide Microarray, supplied by Pepscan Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "A conserved immunogenic and vulnerable site on the coronavirus spike protein delineated by cross-reactive monoclonal antibodies"

Article Title: A conserved immunogenic and vulnerable site on the coronavirus spike protein delineated by cross-reactive monoclonal antibodies

Journal: Nature Communications

doi: 10.1038/s41467-021-21968-w

a Antibody-mediated neutralization of infection of luciferase-encoding VSV particles pseudotyped with spike proteins of MERS-CoV, SARS-CoV, SARS-CoV-2 and HCoV-OC43. Pseudotyped VSV particles pre-incubated with antibodies at indicated concentrations were used to infect VeroCCL81 cells (MERS-S pseudotyped VSV), VeroE6 cells (SARS-S and SARS2-S pseudotyped VSV) or HRT-18 cells (OC43-S pseudotyped VSV) and luciferase activities in cell lysates were determined at 20 h post transduction to calculate infection (%) relative to non-antibody-treated controls. The average ± SD ( n ≥ 6) from at least two independent experiments performed is shown. Iso-CTRL: an anti-Strep-tag human monoclonal antibody was used as an antibody isotype control. The IC 50 and IC 90 values were shown. Source data are provided as a Source Data file. b Antibody-mediated neutralization of MERS-CoV, SARS-CoV and SARS-CoV-2 infection. Neutralization of authentic viruses was performed using a plaque reduction neutralization test (PRNT) on VeroCCL81 cells (MERS-CoV) or VeroE6 (SARS-CoV and SARS-CoV-2) as described earlier  ,  . The experiment was performed with triplicate samples, the average ± SD is shown. The IC 50 and IC 90 values were shown. Source data are provided as a Source Data file. c ELISA-based receptor-binding inhibition assay. MERS-S ecto pre-incubated with serially diluted mAbs was added to ELISA plates coated with soluble human DPP4. The binding of MERS-S ecto to DPP4 was detected using an HRP-conjugated antibody recognizing the C-terminal Strep-tag on MERS-S ecto . Data points represent the average ± SD, for n = 3 replicates from two independent experiments. Source data are provided as a Source Data file. d Cell-cell fusion inhibition assay. Huh-7 cells—transfected with plasmid expressing (GFP-tagged) MERS-CoV S were pre-incubated in the presence or absence of 1.6C7 and 28D9, or an irrelevant iso-type control antibody (Iso-CTRL), and then treated with trypsin to activate the membrane fusion function of the MERS-CoV S protein. The formation of MERS-S mediated syncytia was visualized by fluorescence microscopy. Merged images of MERS-S-GFP expressing cells (green) and DAPI-stained cell nuclei (blue) are shown. The experiment was performed twice, data from a representative experiment is shown. Source data are provided as a Source Data file.
Figure Legend Snippet: a Antibody-mediated neutralization of infection of luciferase-encoding VSV particles pseudotyped with spike proteins of MERS-CoV, SARS-CoV, SARS-CoV-2 and HCoV-OC43. Pseudotyped VSV particles pre-incubated with antibodies at indicated concentrations were used to infect VeroCCL81 cells (MERS-S pseudotyped VSV), VeroE6 cells (SARS-S and SARS2-S pseudotyped VSV) or HRT-18 cells (OC43-S pseudotyped VSV) and luciferase activities in cell lysates were determined at 20 h post transduction to calculate infection (%) relative to non-antibody-treated controls. The average ± SD ( n ≥ 6) from at least two independent experiments performed is shown. Iso-CTRL: an anti-Strep-tag human monoclonal antibody was used as an antibody isotype control. The IC 50 and IC 90 values were shown. Source data are provided as a Source Data file. b Antibody-mediated neutralization of MERS-CoV, SARS-CoV and SARS-CoV-2 infection. Neutralization of authentic viruses was performed using a plaque reduction neutralization test (PRNT) on VeroCCL81 cells (MERS-CoV) or VeroE6 (SARS-CoV and SARS-CoV-2) as described earlier , . The experiment was performed with triplicate samples, the average ± SD is shown. The IC 50 and IC 90 values were shown. Source data are provided as a Source Data file. c ELISA-based receptor-binding inhibition assay. MERS-S ecto pre-incubated with serially diluted mAbs was added to ELISA plates coated with soluble human DPP4. The binding of MERS-S ecto to DPP4 was detected using an HRP-conjugated antibody recognizing the C-terminal Strep-tag on MERS-S ecto . Data points represent the average ± SD, for n = 3 replicates from two independent experiments. Source data are provided as a Source Data file. d Cell-cell fusion inhibition assay. Huh-7 cells—transfected with plasmid expressing (GFP-tagged) MERS-CoV S were pre-incubated in the presence or absence of 1.6C7 and 28D9, or an irrelevant iso-type control antibody (Iso-CTRL), and then treated with trypsin to activate the membrane fusion function of the MERS-CoV S protein. The formation of MERS-S mediated syncytia was visualized by fluorescence microscopy. Merged images of MERS-S-GFP expressing cells (green) and DAPI-stained cell nuclei (blue) are shown. The experiment was performed twice, data from a representative experiment is shown. Source data are provided as a Source Data file.

Techniques Used: Neutralization, Infection, Luciferase, Incubation, Transduction, Strep-tag, Control, Plaque Reduction Neutralization Test, Enzyme-linked Immunosorbent Assay, Binding Assay, Inhibition, Transfection, Plasmid Preparation, Expressing, Membrane, Fluorescence, Microscopy, Staining

a Antibody binding competition analysed by biolayer interferometry. Immobilized MERS-S ecto antigen was saturated in binding with a given mAb (step 1) and then exposed to binding by a second mAb (step 2). Additional binding of the second antibody indicates the presence of an unoccupied epitope, whereas lack of binding indicates epitope blocking by mAb1. As a control, the first mAb was also included in the second step to check for self-competition. The competitive binding was tested for the S2-targeting 1.6C7 and 28D9 antibodies and a MERS-S1 antibody control (7.7G6)  . Source data are provided as a Source Data file. b 1.6C7 and 28D9 recognize a linear epitope. ELISA binding curves of 1.6C7 and 28D9 to untreated MERS-S ecto (non-denatured: ‘nd’) versus MERS-S ecto that was heat-denatured in the presence of SDS and DTT (denatured: ‘d’). Two antibodies targeting the MERS-S1 domain (7.7G6) and the 8-residue long linear Strep-tag epitope (anti-strep) were used as controls. Source data are provided as a Source Data file. c 1.6C7/28D9 epitope maps to a 15-aa long stem region upstream of HR2 in MERS-S. ELISA-reactivity of 1.6C7 and 28D9 to a peptide library of 30-amino acid long peptides (with 15-a.a. overlap) covering the conserved C-terminal part of the MERS-S ecto (residues 869-1,288). Both antibodies reacted with two peptides (blue and orange bars), and with a peptide corresponding to their 15-a.a. long overlap (green bar; MERS-S residues 1,229–1,243). The position of the epitope containing region is indicated in the MERS-S protein schematic with the spike subunits (S1 and S2), S1 domains (A through D), fusion peptide (FP), heptad repeat 1 (HR1), heptad repeat 2 (HR2) and transmembrane domain (TM) annotated. Source data are provided as a Source Data file. d The 1.6C7/28D9 epitope maps to a 8-aa long peptide ‘DELDEFFK’ detected by ELISA. ELISA binding curves of 1.6C7 and 28D9 to N- and C-terminally truncated versions of the 15-mer peptide fragment of MERS-S. Data points represent the average from n = 2 technical replicates. Source data are provided as a Source Data file.
Figure Legend Snippet: a Antibody binding competition analysed by biolayer interferometry. Immobilized MERS-S ecto antigen was saturated in binding with a given mAb (step 1) and then exposed to binding by a second mAb (step 2). Additional binding of the second antibody indicates the presence of an unoccupied epitope, whereas lack of binding indicates epitope blocking by mAb1. As a control, the first mAb was also included in the second step to check for self-competition. The competitive binding was tested for the S2-targeting 1.6C7 and 28D9 antibodies and a MERS-S1 antibody control (7.7G6) . Source data are provided as a Source Data file. b 1.6C7 and 28D9 recognize a linear epitope. ELISA binding curves of 1.6C7 and 28D9 to untreated MERS-S ecto (non-denatured: ‘nd’) versus MERS-S ecto that was heat-denatured in the presence of SDS and DTT (denatured: ‘d’). Two antibodies targeting the MERS-S1 domain (7.7G6) and the 8-residue long linear Strep-tag epitope (anti-strep) were used as controls. Source data are provided as a Source Data file. c 1.6C7/28D9 epitope maps to a 15-aa long stem region upstream of HR2 in MERS-S. ELISA-reactivity of 1.6C7 and 28D9 to a peptide library of 30-amino acid long peptides (with 15-a.a. overlap) covering the conserved C-terminal part of the MERS-S ecto (residues 869-1,288). Both antibodies reacted with two peptides (blue and orange bars), and with a peptide corresponding to their 15-a.a. long overlap (green bar; MERS-S residues 1,229–1,243). The position of the epitope containing region is indicated in the MERS-S protein schematic with the spike subunits (S1 and S2), S1 domains (A through D), fusion peptide (FP), heptad repeat 1 (HR1), heptad repeat 2 (HR2) and transmembrane domain (TM) annotated. Source data are provided as a Source Data file. d The 1.6C7/28D9 epitope maps to a 8-aa long peptide ‘DELDEFFK’ detected by ELISA. ELISA binding curves of 1.6C7 and 28D9 to N- and C-terminally truncated versions of the 15-mer peptide fragment of MERS-S. Data points represent the average from n = 2 technical replicates. Source data are provided as a Source Data file.

Techniques Used: Binding Assay, Blocking Assay, Control, Enzyme-linked Immunosorbent Assay, Residue, Strep-tag

a Schematic representation of the MERS-CoV spike protein. The spike subunits (S1 and S2), S1 domains (A through D), fusion peptide (FP), heptad repeat 1 (HR1), heptad repeat 2 (HR2) and transmembrane domain (TM) are annotated. b Spike protein peptide microarray analysis using MERS-positive human and dromedary camel sera. 905 overlapping peptides covering the entire MERS-CoV S ectodomain (residues 1–1296) were synthesized with an offset of one or two residues. The binding of five convalescent MERS-positive human (H1–H5) and four dromedary camel (D1–D4) sera to the peptide library, as well as a MERS-negative serum from human (H-CTRL) or camel (D-CTRL) was assessed in a PEPSCAN-based ELISA (Lelystad, The Netherlands). A cumulative heatmap of signal intensities for individual peptides are shown. Signal intensities increase from light reddish to red, whereas white corresponds to background signal. Source data are provided as a Source Data file. c Reactivity of the human and dromedary sera to peptides covering the 1.6C7/28D9 epitope region (epitope core sequence highlighted in red). Source data are provided as a Source Data file.
Figure Legend Snippet: a Schematic representation of the MERS-CoV spike protein. The spike subunits (S1 and S2), S1 domains (A through D), fusion peptide (FP), heptad repeat 1 (HR1), heptad repeat 2 (HR2) and transmembrane domain (TM) are annotated. b Spike protein peptide microarray analysis using MERS-positive human and dromedary camel sera. 905 overlapping peptides covering the entire MERS-CoV S ectodomain (residues 1–1296) were synthesized with an offset of one or two residues. The binding of five convalescent MERS-positive human (H1–H5) and four dromedary camel (D1–D4) sera to the peptide library, as well as a MERS-negative serum from human (H-CTRL) or camel (D-CTRL) was assessed in a PEPSCAN-based ELISA (Lelystad, The Netherlands). A cumulative heatmap of signal intensities for individual peptides are shown. Signal intensities increase from light reddish to red, whereas white corresponds to background signal. Source data are provided as a Source Data file. c Reactivity of the human and dromedary sera to peptides covering the 1.6C7/28D9 epitope region (epitope core sequence highlighted in red). Source data are provided as a Source Data file.

Techniques Used: Peptide Microarray, Synthesized, Binding Assay, Enzyme-linked Immunosorbent Assay, Sequencing



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Image Search Results


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).

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

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).

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

A Schematic model showing the PTK PamChip peptide microarray system and experimental design. B Heat map of up-and down-regulated Tyrosine (PTK assay) bait peptide phosphorylation of lorlatinib (4 h) versus DMSO treatment by peptide chip array ( n = 3 biological replicates) in H3122 and H2228 cell lines. Significant peptides ( p < 0.05) were clustered using hierarchical order and Euclidean/ward algorithm. C , D Volcano plots of the bait peptide phosphorylation from PTK chip arrays in A showing significant up- (Red) and downregulation (blue) of peptide phosphorylation as defined by ANOVA and post-hoc Dunnett’s test versus DMSO treatment ( p < 0.05). Non-significant peptides were highlighted in grey. Downregulated peptides were highlighted in light blue. ERBB family and AKT1 phosphopeptides were highlighted in green and black bold colours, respectively. E Venn diagram was generated to compare the phosphorylated Tyr peptides (up- and downregulated) after a 4 h treatment with lorlatinib in H3122 and H2228 cell lines. Tables of the specific phosphorylated Tyr sites and the associated proteins are shown. The down-regulated proteins were highlighted in cyan colour. The upregulated proteins selected for further analysis were highlighted in bold with green and black colours.

Journal: Cell Death & Disease

Article Title: Targeting ERBB3 and AKT to overcome adaptive resistance in EML4-ALK-driven non-small cell lung cancer

doi: 10.1038/s41419-024-07272-7

Figure Lengend Snippet: A Schematic model showing the PTK PamChip peptide microarray system and experimental design. B Heat map of up-and down-regulated Tyrosine (PTK assay) bait peptide phosphorylation of lorlatinib (4 h) versus DMSO treatment by peptide chip array ( n = 3 biological replicates) in H3122 and H2228 cell lines. Significant peptides ( p < 0.05) were clustered using hierarchical order and Euclidean/ward algorithm. C , D Volcano plots of the bait peptide phosphorylation from PTK chip arrays in A showing significant up- (Red) and downregulation (blue) of peptide phosphorylation as defined by ANOVA and post-hoc Dunnett’s test versus DMSO treatment ( p < 0.05). Non-significant peptides were highlighted in grey. Downregulated peptides were highlighted in light blue. ERBB family and AKT1 phosphopeptides were highlighted in green and black bold colours, respectively. E Venn diagram was generated to compare the phosphorylated Tyr peptides (up- and downregulated) after a 4 h treatment with lorlatinib in H3122 and H2228 cell lines. Tables of the specific phosphorylated Tyr sites and the associated proteins are shown. The down-regulated proteins were highlighted in cyan colour. The upregulated proteins selected for further analysis were highlighted in bold with green and black colours.

Article Snippet: To identify adaptive response mechanisms, we examined the landscape of tyrosine phosphorylation upon acute LOR treatment in EML4-ALK-positive NSCLC cell lines using PamGene microarray peptide assay, known as protein tyrosine kinase (PTK) (Fig. ).

Techniques: Peptide Microarray, Generated