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Reactivity of commercial antibodies targeting human histone H2B N-terminal tail on the Intel array platform

ABSTRACT: The aim of the study was to test the Intel array platform and map the H2B epitopes bound by commercial antibody reagents. We also tested the ability of SET7 PKMT to methylate Intel array peptides. We diluted antibody reagents 1:1000 and incubated dilutions on the silicon-based Intel array platorm. We mapped antibody binding epitopes and SET7 PKMT methylation target peptides using a novel, silicon-based array platform with 8820 individual peptide features comprising every possible contiguous peptide from 1-21 a.a. in length within the 21-mer N-terminal tail of human histone H2B.

ORGANISM(S): Oryctolagus cuniculus

SUBMITTER: Jordan Price

PROVIDER: E-GEOD-34034 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Reactivity of commercial antibodies targeting human histone H2B N-terminal tail on the Intel array platform

Project description:The aim of the study was to test the Intel array platform and map the H2B epitopes bound by commercial antibody reagents. We also tested the ability of SET7 PKMT to methylate Intel array peptides. We diluted antibody reagents 1:1000 and incubated dilutions on the silicon-based Intel array platorm.

2011-12-02 | GSE34034 | GEO

Influenza peptide reactivity within a cohort of age-stratified individuals pre and post vaccination with the 2008/2009 trivalent influenza vaccine

Project description:The aim of the study was to test the hypothesis that peptide array reactivity will reveal patterns that can be associated with poor or effective neutralization response to seasonal influenza vaccine We diluted patient sera 1:80 and incubated dilutions on a streptavidin-coated glass surface that was printed with biotinylated peptides derived from seasonal strains of influenza hemagglutinin included in the 2008/2009 trivalent influenza vaccine In this study, serum from 76 study participants, subsetted into young (?30 years old) and elderly (?60 years old) categories were profiled for IgG/IgM antibody reactivity to influenza hemagglutinin peptudes. Using multivariate linear regression and correcting for multiple hypothesis testing by cross-validation, ee found that patterns of peptide reactivity associated with a poor or effective neutralization response to H1N1 and B strains of influenza.

2013-05-01 | E-GEOD-43446 | biostudies-arrayexpress

The Trypanosoma cruzi Antigen and Epitope Atlas: fine characterization of antibody specificities in human Chagas Disease patients across human populations

Project description:These assays represent antibody-binding assays to analyze seroprevalence of antigens and epitopes at the individual level and also to perform fine epitope mapping characterizations. In this screening 392,299 peptides derived from reactive (antigenic) regions of Trypanosoma cruzi proteins were displayed in the form of short peptides (tiling array, overlapped) and assayed with individual serum samples (antibodies) from Chagas Disease patients across the Americas. Sectorized peptide arrays (QX12-plex slides, Roche Nimblegen) were incubated with serum samples (primary antibodies), washed, and then incubated with a fluorescently-labeled anti-human IgG commercial antibody (secondary antibodies). Raw readouts of fluoresence (signal), as well as normalized signal values are provided in this submission for all samples analyzed. All samples were analyzed in duplicate (r1 = replicate 1; r2 = replicate 2).

2022-08-20 | E-MTAB-11655 | biostudies-arrayexpress

PAD4 Inhibition is Sufficient for the Amelioration of Collagen-Induced Arthritis

Project description:To examine the effects of GSK199 treatment on antibody responses, a synovial antigen array analysis was performed using day 35 sera derived from mice with CIA treated with 30 mg/kg qd, 30 mg/kg bid GSK199 or saline. The synovial antigen arrays contain nearly 200 peptides and proteins, both citrullinated and uncitrullinated, representing candidate antigens in RA. Significance Analysis of Microarrays identified 7 and 14 autoantibody reactivities that were statistically decreased in CIA mice treated with 30 mg/kg qd or 30 mg/kg bid GSK199 respectively, compared to PBS treatment. By comparison, over 150 native and citrullinated epitopes on the arrays did not exhibit significant differences in reactivity. Sera from mice treated with 30 mg/kg GSK199 qd exhibited decreased IgG reactivity to native epitopes such as peptides from human fibrinogen alpha and beta chains and keratin while sera from mice treated with 30 mg/kg bid GSK199 demonstrated decreased IgG reactivity to native epitopes including collagen IV, heat shock protein 65, peptides from human fibrinogen alpha and beta chains (two of which match those that were decreased in the 30 mg/kg qd group), and vimentin as compared to saline controls (q < 0.1%). Decreased autoantibody reactivity to citrullinated peptides from vimentin and fibromodulin were also observed in sera from mice treated with 30 mg/kg bid GSK199 compared to controls treated with saline (q < 0.1%), but not by GSK199 30 mg/kg qd. There were no differences between treatment with GSK199 or saline alone in the levels of autoantibodies to 38 other citrullinated proteins in the array. These data demonstrated that GSK199 treatment decreases the development of autoantibodies in CIA and impairs epitope spreading to a degree similar to that observed with pan-PAD inhibition.

2015-10-08 | GSE69680 | GEO

Autoimmune peptide antigen IgG and IgM reactivity in B6 and BXD2 mice with spontaneous systemic autoimmune disease

Project description:The aim of the study was to determine B6 and BXD2 mouse sera IgG and IgM reactivity to linear peptide epitopes at different ages. Serum from B6 or BXD2 mice was diluted at 1:200 for IgG-specific analysis or 1:1000 for IgM specific analysis and incubated on a PEPperPRINT peptide microarray platform printed with peptide autoantigens. 80 pre-selected peptides based on a previous screen of pooled mouse serum BXD2 against the PEPperCHIPÂ® Autoimmunity Microarray with 2,733 linear B-cell epitopes were printed in duplicate in 16 copies on a custom PEPperCHIPÂ® Peptide Microarray. Flag (DYKDDDDKGG) and HA (YPYDVPDYAG) control peptides (10 spots each control) were randomly distributed in each array copy as controls. Sera from 2, 5, or 9 month old B6 or BXD2 mice was profiled for anti-mouse IgG or IgM-specific analysis of autoantibody reactivity to the peptide auto-epitopes.

2015-02-28 | E-GEOD-65234 | biostudies-arrayexpress

Epitope mapping of a monoclonal antibody directed against Neisserial Heparin Binding Antigen by peptide microarray

Project description:The aim of the study was to determine the epitope targeted by 31E10/E7 mouse monoclonal antibody (mAb) and the cross-reactivity to linear peptide epitopes of 10 different Neisserial Heparin Binding Antigen (NHBA) variants. mAb 31E10/E7 was diluted at 1:200 and incubated on a custom PepStar Peptide Microarray platform printed with 560 different peptides in three independent experiments.

2016-05-27 | GSE81377 | GEO

In vitro expression in FGFR inhibitor sensitive and resistant cell lines

Project description:The challenge of developing effective pharmacodynamic biomarkers for pre-clinical and clinical testing of FGFR signalling inhibition is significant. Assays that rely on the measurement of phospho-protein epitopes can be limited by the availability of effective antibody detection reagents. Transcript profiling enables accurate quantification of many biomarkers and a broader representation of pathway modulation. To identify dynamic transcript biomarkers of FGFR signalling inhibition by AZD4547, a potent inhibitor of FGF receptor 1, 2 and 3, a gene expression profiling study was performed in FGFR2 amplified drug sensitive tumour cell lines.

2016-08-25 | E-MTAB-4749 | biostudies-arrayexpress

Immunogenic epitopes of the meningioma-associated antigen TXNDC16

Project description:We used customized peptide arrays of 163 TXNDC16 peptides to identify immunogenic TXNDC16 epitopes and asked whether discrimination of meningioma and control sera is possible with a specific subset selection of all immunogenic epitopes. CelluSpotsM-bM-^DM-" peptide arrays were manufactured by Intavis (Germany) with 163 TXNDC16 spanning peptides spotted as two replicate subarrays. 15-mer peptides overlapping by 10 amino acids were covalently bound to cellulose membranes with their C-termini. Please note that the non_normalized.txt contains Ch1 Mean values for all samples (each sequence represented in duplicates) and the identifiers in the 'index' column corresponds to those in the 'index' column in raw gpr files; sample data table contains classification values (for each sequence) described in the sample data processing field.

2014-07-01 | E-GEOD-58949 | biostudies-arrayexpress

Immunogenic epitopes of the meningioma-associated antigen TXNDC16

2014-07-01 | GSE58949 | GEO

Proteomic profiling of paired interstitial fluids revealed dysregulated pathways and salivary NID1 as a biomarker of oral cavity squamous cell carcinoma

Project description:Patients with oral cavity squamous cell carcinoma (OSCC) are frequently first diagnosed at an advanced stage, leading to poor prognosis and high mortality rate. Early detection of OSCC using body fluid-accessible biomarkers may help improve the prognosis and survival rate of OSCC patients. As tumor interstitial fluid (TIF) is a proximal fluid enriched with cancer-related proteins, it is therefore a valuable reservoir suitable for the discovery of cancer biomarkers as well as dysregulated biological pathways in tumor microenvironment. Thus, we harvested and analyzed the paired tumor (TIF) and adjacent noncancerous (NIF) interstitial fluids from ten OSCC patients using one-dimensional gel electrophoresis coupled with the nano-liquid chromatography-tandem mass spectrometry (GeLC-MS/MS) approach and label-free spectral counting method. The results showed that 113 proteins were up-regulated in at least six OSCC TIFs compared to their respective NIFs, and that the aminoacyl tRNA biosynthesis was statistically enriched in TIFs by Gene Set Enrichment Analysis (GSEA). The enrichment of aminoacyl tRNA biosynthesis in OSCC was verified that four members (IARS, KARS, WARS, and YARS) were revealed to be elevated in OSCC by IHC (n = 12). Among the 113 up-regulated proteins, two candidates (NID1 and SERPINH1) were selected based on an integrated bioinformatics analysis and verified that the salivary NID1 levels were significantly higher in OSCC patients (n = 48) compared to health (n = 51) and OPMD individuals (n = 53). Moreover, the IHC results (n = 222) showed that the expression of NID1 was higher in OSCC compared to adjacent noncancerous epithelium and correlated with pathological T, N, and overall stage as well as survivals of OSCC patients. These results collectively indicate that the analysis of TIF is help to understand the tumor microenvironment, and that salivary NID1 is a potentially useful biomarker for the OSCC.

2019-08-15 | PXD008654 | Pride

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