Journal: Virology Journal

Article Title: Development and characterization of serotype-specific monoclonal antibodies against the dengue virus-4 (DENV-4) non-structural protein (NS1)

doi: 10.1186/s12985-018-0925-7

Figure Lengend Snippet: SUMO*-DENV-4 NS1 fusion gene construct. Schematic showing the DENV-4 fusion construct within the SUMO* cloning vector used to express recombinant protein in Spodoptera frugiperda (Sf)21 cells

Article Snippet: We determined the serotype-specificity of the above described NS1 capture ELISA using culture supernatants obtained from DENV-infected Vero cells and commercially available rNS1 of flaviviruses, including all four DENV serotypes and expressed in mammalian cell line (NativeAntigen, Oxfordshire, UK).

Techniques: Construct, Clone Assay, Plasmid Preparation, Recombinant

Journal: Virology Journal

Article Title: Development and characterization of serotype-specific monoclonal antibodies against the dengue virus-4 (DENV-4) non-structural protein (NS1)

doi: 10.1186/s12985-018-0925-7

Figure Lengend Snippet: SUMO*-NS1 fusion protein expression analysis. a Coomassie-blue stained 12% SDS-PAGE gel. Lane 1: SUMO*-DENV-4 NS1 under non-reducing conditions (dimer); Lane 2: SUMO*-DENV-4 NS1 under reducing conditions (monomer); Lane 3: MW of markers in kDa. b Western blot of SUMO*-DENV-4 NS1 protein using anti-HIS antibody: Lane 1: Molecular Marker; Lane 2: secreted NS1; Lane 3: cell lysate of solubilized of SUMO*- DENV-4 NS1

Article Snippet: We determined the serotype-specificity of the above described NS1 capture ELISA using culture supernatants obtained from DENV-infected Vero cells and commercially available rNS1 of flaviviruses, including all four DENV serotypes and expressed in mammalian cell line (NativeAntigen, Oxfordshire, UK).

Techniques: Expressing, Staining, SDS Page, Western Blot, Marker

Journal: Virology Journal

Article Title: Development and characterization of serotype-specific monoclonal antibodies against the dengue virus-4 (DENV-4) non-structural protein (NS1)

doi: 10.1186/s12985-018-0925-7

Figure Lengend Snippet: Anti-NS1 monoclonal antibodies isotype, epitope and reactivity to DENV1–4 NS1. Monoclonal antibody reactivity to native DENV1–4 NS1 in the fixed cell ELISA and iELISA

Article Snippet: We determined the serotype-specificity of the above described NS1 capture ELISA using culture supernatants obtained from DENV-infected Vero cells and commercially available rNS1 of flaviviruses, including all four DENV serotypes and expressed in mammalian cell line (NativeAntigen, Oxfordshire, UK).

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Cell Culture

Journal: Virology Journal

Article Title: Development and characterization of serotype-specific monoclonal antibodies against the dengue virus-4 (DENV-4) non-structural protein (NS1)

doi: 10.1186/s12985-018-0925-7

Figure Lengend Snippet: Anti-NS1 Monoclonal Antibodies (MAbs) reactivity to SUMO*-NS1 fusion protein. Western blot assay to determine the reactivity of hybridoma supernatants to SUMO*-NS1 fusion protein as well as unfused commercial rNS1 protein expressed in mammalian cell,; M; molecular markers; Lanes 1 and 2: 3H7A9; Lanes 3 and 4: 4B6C10; Lanes 5 and 6: 6D4B10; Lanes 7 and 8: 8A6F2; Lanes 9 and 10: 10H8F7; Lanes 11 and 12: 10H10B5; Lanes 13 and 14: Antiserum of mouse ME 1:100, reactivity to SUMO*-DENV NS1 fusion protein and unfused commercial DENV rNS1 expressed in mammalian cell, respectively. The sizes of the monomer and dimer for SUMO-rNS1 are ~ 62 kDa and 120 kDa due ~ 15 kDa SUMO tag whereas the sizes for monomer and dimer of unfused commercial rNS1 (NativeAntigen) are 48 kDa and 80 kDa

Article Snippet: We determined the serotype-specificity of the above described NS1 capture ELISA using culture supernatants obtained from DENV-infected Vero cells and commercially available rNS1 of flaviviruses, including all four DENV serotypes and expressed in mammalian cell line (NativeAntigen, Oxfordshire, UK).

Techniques: Western Blot

Journal: Virology Journal

Article Title: Development and characterization of serotype-specific monoclonal antibodies against the dengue virus-4 (DENV-4) non-structural protein (NS1)

doi: 10.1186/s12985-018-0925-7

Figure Lengend Snippet: Serotype-specificity of monoclonal antibodies (MAbs 3H7A9, 8A6F2, 6D4B10 and 10H10B5) against DENV-4 determined by iELISA. The bars show mean optical density (OD) at 450 nm which measures MAb reactivity to NS1s to all four DENV-4 serotypes, yellow fever virus (YFV), West Nile virus (WNV) and SUMO protein. MAbs 3H7A9, 8A6F2, and 6D4B10 represent anti-NS1 MAbs specific to DENV-4. MAb 10H10B5, which is reactive to SUMO*, represents one of the three non-specific MAbs generated from immunization of SUMO*-DENV-4 rNS1 fusion protein. The OD value for DENV-4-serotype specific MAbs against rNS1 of other flaviviruses and recombinant SUMO protein was < 0. By contrary, MAbs reactive only to SUMO* protein (10H10B5) did not show any reactivity to rNS1 to YFV and WNV

Article Snippet: We determined the serotype-specificity of the above described NS1 capture ELISA using culture supernatants obtained from DENV-infected Vero cells and commercially available rNS1 of flaviviruses, including all four DENV serotypes and expressed in mammalian cell line (NativeAntigen, Oxfordshire, UK).

Techniques: Virus, Generated, Recombinant

Journal: Virology Journal

Article Title: Development and characterization of serotype-specific monoclonal antibodies against the dengue virus-4 (DENV-4) non-structural protein (NS1)

doi: 10.1186/s12985-018-0925-7

Figure Lengend Snippet: DENV-4 serotype-specificity MAbs 3H7A9, 8A6F2, 6D4B10 and 10H10B5 as determined by fixed cell ELISA. The bars showed mean optical density at 450 nm that measures MAb’s reactivity to dimeric NS1 expressed on DENV1–4-infected Vero cells. Control included mock-infected Vero cells

Article Snippet: We determined the serotype-specificity of the above described NS1 capture ELISA using culture supernatants obtained from DENV-infected Vero cells and commercially available rNS1 of flaviviruses, including all four DENV serotypes and expressed in mammalian cell line (NativeAntigen, Oxfordshire, UK).

Techniques: Enzyme-linked Immunosorbent Assay, Infection

Journal: Virology Journal

Article Title: Development and characterization of serotype-specific monoclonal antibodies against the dengue virus-4 (DENV-4) non-structural protein (NS1)

doi: 10.1186/s12985-018-0925-7

Figure Lengend Snippet: Anti-NS1 monoclonal antibodies isotype, epitope and reactivity to DENV1–4 NS1. Isotype, epitope type and iELISA results of six anti-NS1 DENV-4 monoclonal antibodies

Article Snippet: We determined the serotype-specificity of the above described NS1 capture ELISA using culture supernatants obtained from DENV-infected Vero cells and commercially available rNS1 of flaviviruses, including all four DENV serotypes and expressed in mammalian cell line (NativeAntigen, Oxfordshire, UK).

Techniques: Recombinant

Journal: Virology Journal

Article Title: Development and characterization of serotype-specific monoclonal antibodies against the dengue virus-4 (DENV-4) non-structural protein (NS1)

doi: 10.1186/s12985-018-0925-7

Figure Lengend Snippet: Analysis of optimal capture/detector MAb pair for development of DENV-4 serotype specific NS1 capture ELISA. The curves represent limit of detection (LOD) curve for matched pairs: a 8A6F2 and biotinylated 6D4B10, and b 6D4B10 and biotinylated 8A6F2. The optical density (OD) values at 450 nm were obtained at various concentration of DENV-4 rNS1 with an optimal concentrations for coating antibody, 5-10 μg/ml; capture antibody, 1:2000 dilution and streptavidin-tagged horseradish peroxidase (HRP-SP), 1:2000. WNV was used as the control

Article Snippet: We determined the serotype-specificity of the above described NS1 capture ELISA using culture supernatants obtained from DENV-infected Vero cells and commercially available rNS1 of flaviviruses, including all four DENV serotypes and expressed in mammalian cell line (NativeAntigen, Oxfordshire, UK).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Nature Communications

Article Title: NETosis and thrombosis in vaccine-induced immune thrombotic thrombocytopenia

doi: 10.1038/s41467-022-32946-1

Figure Lengend Snippet: a PF4 and PF4-heparin ELISA experiment of VITT serum and controls. The cut-off, 0.50 OD units (Vax, VTE n = 7; AB, VITT Pt and HIT n = 3). b 14 C-serotonin release assay for VITT samples with buffer alone, PF4 (10 μg/mL), 0.1 or 100 U/mL heparin or IV.3 antibody (50 μg/mL). Each dot represents the mean of assays done in triplicate. The cut-off was set at 20% CPM. c Platelet aggregation responses. Purified IgG from VITT patients induced aggregation in platelet-rich plasma (red, blue and black traces). Blockage of FcγRIIa with IV.3 inhibited aggregation (purple, green and light blue traces). d Nucleosomal CitH3 (H3R8Cit ELISA, * p = 0.03), e myeloperoxidase (ELISA, ** p = 0.01), and f cfDNA (PicoGreen fluorescence assay, **** p < 0.0001) levels in VITT patients’ plasma ( n = 7) relative to controls ( n = 7) were determined. g Representative side and forward scatter flow cytometry plot backgated for neutrophils (yellow) and monocytes (blue) from VITT patient’s and vaccine control blood. LDG are indicated. h LDG quantitated as number of LDG events relative to 200 monocytes ( n = 7, except VITT n = 3; * p = 0.02). i Representative plot of NPA from VITT and vaccine control blood. j Quantification of NPA in VITT ( n = 7, except VITT n = 3; ** p = 0.004). k Representative plot of NETs from VITT and vaccine control blood. Quantification of NETs in VITT in l whole blood ( n = 7, except VITT n = 3; ** p = 0.005) and in m LDG population ( n = 6, except VITT n = 3; * p = 0.02). MPO + , CitH3 + double positive cells within the CD15 + population were defined as neutrophils undergoing NETosis. The percentage of gated events is indicated in each quadrant. Statistics: Kruskal-–Wallis ANOVA with Dunn’s correction. Data are presented as a , j , l , m mean ± SD; h mean ± SEM. OD optical density units, CPM counts per minute, Vax ctrl healthy vaccinated subject IgG; VTE venous thromboembolism patient IgG, ICU intensive care unit patient IgG, NPA neutrophil-platelet aggregates, LDG low density granulocytes, cfDNA cell-free DNA, CitH3 citrullinated histone H3, Pt patient. Source data are provided in the Source Data file.

Article Snippet: Plasma levels of myeloperoxidase and citrullinated histone H3 were determined using the human myeloperoxidase ELISA kit (ab119605, Abcam) and H3R8Cit ELISA Capture and Detection kit (R&D143002, EpiCypher) , respectively, following the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Release Assay, Purification, Clinical Proteomics, Fluorescence, Flow Cytometry, Control

Journal: medRxiv

Article Title: Performance Characteristics of Six Immunoglobulin M (IgM) ELISA Assays Used for Laboratory Confirmation of Measles

doi: 10.1101/2022.09.02.22279538

Figure Lengend Snippet: (A) Absorbance values for samples tested by the CDC IgM ELISA for each commercial kit. The absorbance values of the positive (measles NP antigen) and negative (SF9 control antigen) are shown for each sample used on the comparison for each commercial ELISA test. The cut-off criteria for the negative and positive results are shown by the orange line (P-N=0.1) and the black line (P/N=3.0). Black circles indicate CDC IgM results that qualitatively agree with the indicated commercial test. Red squares indicate false positive results. Blue triangles indicate false negative results that were confirmed as positive by PCR. (B) Positive and Negative cut-off values for samples tested by the CDC IgM ELISA in comparison for each commercial kit. The cut-off criteria for P/N vs. P-N values are shown by dotted lines. Black circles indicate CDC IgM results that qualitatively agree with the indicated commercial test. Red squares indicate false positive results. Blue triangles indicate false negative results that were confirmed as positive by PCR.

Article Snippet: The five measles IgM ELISA assay kits that were compared included: Euroimmun Anti-Measles Virus NP IgM ELISA, (Euroimmun, Lübeck Germany catalog no. EL2610-9601-4M), Virion/Serion ELISA classic measles virus IgM (Virion/Serion, Würzburg Germany catalog no. ESR102M), Awareness Technology (AwareTech) ReQuest Measles IgM (Quest International Inc., Doral, FL catalog no. 01-190M), IBL International Measles Virus IgM micro-capture ELISA (IBL International, Hambürg Germany catalog no. RE57151), and Trinity Biotech Captia Measles IgM ELISA, (Trinity Biotech, Jamestown NY catalog no. K140455).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Comparison

Journal: medRxiv

Article Title: Performance Characteristics of Six Immunoglobulin M (IgM) ELISA Assays Used for Laboratory Confirmation of Measles

doi: 10.1101/2022.09.02.22279538

Figure Lengend Snippet: (A) Cumulative false negative and false positive results from each commercial ELISA were calculated based on agreement with the CDC IgM ELISA. The false results are expressed as a percentage of total samples tested. (B) The fraction of false negative results from each commercial kit are shown for cases that were IgM-positive by the CDC ELISA and confirmed by RT-PCR in accompanying throat swabs.

Article Snippet: The five measles IgM ELISA assay kits that were compared included: Euroimmun Anti-Measles Virus NP IgM ELISA, (Euroimmun, Lübeck Germany catalog no. EL2610-9601-4M), Virion/Serion ELISA classic measles virus IgM (Virion/Serion, Würzburg Germany catalog no. ESR102M), Awareness Technology (AwareTech) ReQuest Measles IgM (Quest International Inc., Doral, FL catalog no. 01-190M), IBL International Measles Virus IgM micro-capture ELISA (IBL International, Hambürg Germany catalog no. RE57151), and Trinity Biotech Captia Measles IgM ELISA, (Trinity Biotech, Jamestown NY catalog no. K140455).

Techniques: Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction

Journal: Tropical Medicine and Infectious Disease

Article Title: Development of an Anti-Zika and Anti-Dengue IgM ELISA Assay: Evaluation of Cross Reactivity and Validation

doi: 10.3390/tropicalmed7110348

Figure Lengend Snippet: Isotypes of ZIKV 8-8-11 and DENV 8G2-12-21 mAbs.

Article Snippet: Comparison with other commercially available assays A total of 159 human serum samples were analysed by both the CHORUS Zika IgM capture assay and Zika virus IgM micro-capture ELISA (IBL international).

Techniques:

Journal: Tropical Medicine and Infectious Disease

Article Title: Development of an Anti-Zika and Anti-Dengue IgM ELISA Assay: Evaluation of Cross Reactivity and Validation

doi: 10.3390/tropicalmed7110348

Figure Lengend Snippet: Specificity of the ZIKV 8-8-11 and DENV 8G2-12-21 mAbs.

Article Snippet: Comparison with other commercially available assays A total of 159 human serum samples were analysed by both the CHORUS Zika IgM capture assay and Zika virus IgM micro-capture ELISA (IBL international).

Techniques:

Journal: Tropical Medicine and Infectious Disease

Article Title: Development of an Anti-Zika and Anti-Dengue IgM ELISA Assay: Evaluation of Cross Reactivity and Validation

doi: 10.3390/tropicalmed7110348

Figure Lengend Snippet: Specificity of the CHORUS Zika and Dengue IgM Capture assays.

Article Snippet: Comparison with other commercially available assays A total of 159 human serum samples were analysed by both the CHORUS Zika IgM capture assay and Zika virus IgM micro-capture ELISA (IBL international).

Techniques:

Journal: Tropical Medicine and Infectious Disease

Article Title: Development of an Anti-Zika and Anti-Dengue IgM ELISA Assay: Evaluation of Cross Reactivity and Validation

doi: 10.3390/tropicalmed7110348

Figure Lengend Snippet: Comparison between the CHORUS and IBL IgM capture assays.

Article Snippet: Comparison with other commercially available assays A total of 159 human serum samples were analysed by both the CHORUS Zika IgM capture assay and Zika virus IgM micro-capture ELISA (IBL international).

Techniques: Comparison, Virus, Enzyme-linked Immunosorbent Assay

Journal: Tropical Medicine and Infectious Disease

Article Title: Development of an Anti-Zika and Anti-Dengue IgM ELISA Assay: Evaluation of Cross Reactivity and Validation

doi: 10.3390/tropicalmed7110348

Figure Lengend Snippet: Comparison between the CHORUS and Panbio Dengue IgM Capture ELISA.

Article Snippet: Comparison with other commercially available assays A total of 159 human serum samples were analysed by both the CHORUS Zika IgM capture assay and Zika virus IgM micro-capture ELISA (IBL international).

Techniques: Comparison, Enzyme-linked Immunosorbent Assay

Journal: Tropical Medicine and Infectious Disease

Article Title: Development of an Anti-Zika and Anti-Dengue IgM ELISA Assay: Evaluation of Cross Reactivity and Validation

doi: 10.3390/tropicalmed7110348

Figure Lengend Snippet: Analysis of discordant samples with Euroimmun anti-Dengue Elisa (IgM).

Article Snippet: Comparison with other commercially available assays A total of 159 human serum samples were analysed by both the CHORUS Zika IgM capture assay and Zika virus IgM micro-capture ELISA (IBL international).

Techniques: Enzyme-linked Immunosorbent Assay, Virus

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Monophosphoryl lipid A plus IFNγ maturation of dendritic cells induces antigen-specific CD8 + cytotoxic T cells with high cytolytic potential

doi: 10.1007/s00262-010-0843-z

Figure Lengend Snippet: The superior induction of CTLs by MPLA/IFNγ DCs compared to CC-DCs is dependent on IL-12 and not on IL-6 or IL-23. a Blocking antibodies specific for IL-12p40 and IL-6, and an irrelevant antibody were added to MPLA/IFNγ DC-CTL cocultures. 7 days after the second stimulation of the CTLs with DCs (day 17), the CTLs were stained with the MART-1 tetramer (TM) and CD8-FITC. A graph showing the % MART-1 TM cells for all separate cultures of one experiment is shown. The condition treated with anti-IL-12 significantly differed (one-way ANOVA, P < 0.05) from untreated or isotype control treated conditions. A representative experiment out of four is depicted. b Blocking antibodies specific for IL-12p35 and IL-23p19, and an irrelevant antibody were added to MPLA/IFNγ DC-CTL cocultures. 7 days after the third stimulation of the CTLs, the CTLs were stained with the MART-1 TM and CD8-FITC. A graph showing the % MART-1 TM cells for all separate cultures of one experiment is shown. The condition treated with anti-IL-12p35 significantly differed (one-way ANOVA, P < 0.01) from isotype control treated condition

Article Snippet: The following neutralizing antibodies were used; anti-IL-12p40 (C8.6), anti-IL-12p35 (B-T21, Diaclone), anti-IL-23p19 (B-Z23, Diaclone), anti-IL-6 (IL6.8 [ 26 ]).

Techniques: Blocking Assay, Staining, Control

Journal: Vaccines

Article Title: Evaluating the Compatibility of Three Aluminum Salt-Adjuvanted Recombinant Protein Antigens (Trivalent NRRV) Combined with a Mock Trivalent Sabin-IPV Vaccine: Analytical and Formulation Challenges

doi: 10.3390/vaccines12101102

Figure Lengend Snippet: Stability profiles of AH-adsorbed t-sIPV antigens (types 1, 2, and 3) in the presence and absence of different antimicrobial preservatives (APs) as measured by Sabin D-antigen competitive ELISAs. The relative antigen–antibody binding of each AH-adsorbed sIPV antigen to an antigen-specific antibody is shown as a percentage concentration relative to time zero values after storage at 2–8 °C and 15 °C for 3 months, and 37 °C for 1 day, for sIPV1 (Panels ( A , D , G )), sIPV2 (Panels ( B , E , H )), and sIPV3 (Panels ( C , F , I )) in the presence of the indicated AP. TH—thimerosal, 2-PE—2-phenoxy ethanol, PH—phenol, CB—chlorobutanol, MC—m-cresol, BA—benzyl alcohol, MP—methyl paraben, and PP—propyl paraben. Data are presented as the mean ± SD (n = 4).

Article Snippet: Capture and detection antibodies used in the competition enzyme-linked immunoassays (ELISAs), system suitability controls, and reference standards were purchased from Batavia Biosciences (The Netherlands) for three sIPV antigens and Precision Antibody (Columbia, MD, USA) for the three NRRV antigens.

Techniques: Binding Assay, Concentration Assay