Journal: Analytical Chemistry

Article Title: A Novel Quantitative Multi-Component Serological Assay for SARS-CoV-2 Vaccine Evaluation

doi: 10.1021/acs.analchem.1c05264

Figure Lengend Snippet: LLOD of the microarray tests. Upper panel: Calculated MFI values (after blank subtraction) for (left to right) IgG, IgA, and IgM signals of the commercial, pre-pandemic, naïve samples (eight independent experiments) on S2P (orange), RBD (blue), and NC (green). The average (AV), standard deviation (STD), and lower limit of detection (LLOD) for each antibody isotope for each antigen are indicated on the graph (bold black lines and dashed colored lines, respectively), and the values are presented in the lower panel. LLOD values are indicated in bold. No LLOD was determined for IgM-NC, since no acceptance range was determined for this test (due to a high CV value observed during acceptance range determination, Figure S2 ).

Article Snippet: SARS-CoV-2 antigens including S2P, RBD, and NC (1 mg/mL) were spotted separately, as single 300 pL drops, in 18 repeats on 16-pad (16 sub-arrays) nitrocellulose-coated slides (Grace Bio Labs, GBL, Bend, OR) using a non-contact Piezo dispensing microarray spotter (Scienion Inc., Berlin, Germany).

Techniques: Microarray, Standard Deviation

Journal: Ophthalmic research

Article Title: Effect of ischemia duration on autoantibody response in rats undergoing retinal ischemia-reperfusion.

doi: 10.1159/000335965

Figure Lengend Snippet: Fig. 2. Exemplary protein microarray sections incubated with a control serum sample ( a ) or samples of animals after 30 min ( b ), 45 min ( c ), and 90 min ( d ) of ischemia. Antigens were all spotted in triplicates. The antibody reactivities against cyclophilin (I 1), -A-crystallin (I 4), HSP 60 (II 3), GST (II 5), GAPDH (III 1), and SOD (IV 3) are displayed.

Article Snippet: Antigen spotting was performed with a noncontact microarray spotter (sciFLEXARRAYER S3; Scienion, Berlin, Germany).

Techniques: Microarray, Incubation, Control

Journal: Viruses

Article Title: Multiplex Serology for Sensitive and Specific Flavivirus IgG Detection: Addition of Envelope Protein Domain III to NS1 Increases Sensitivity for Tick-Borne Encephalitis Virus IgG Detection

doi: 10.3390/v16020286

Figure Lengend Snippet: Production of orthoflavivirus recombinant NS1 for sensitive and specific microarray detection of orthoflavivirus IgG. ( a ) Recombinant NS1 and ( b ) recombinant EDIII of TBEV, West Nile virus (WNV) lineage 2, dengue virus (DENV) DENV 1 , DENV 2 , DENV 3 , DENV 4 , Zika virus (ZIKV), Japanese encephalitis virus (JEV) and Usutu virus (USUV) were purified and then analyzed via SDS-PAGE.

Article Snippet: TBEV rEDIII and the rNS1 of DENV 1 , DENV 2 , DENV 3 , DENV 4 , JEV, TBEV, USUV, WNV lineage 2 and ZIKV at concentrations of 1.5 mg/mL were mixed with 2× protein printing buffer (GVS, Sandford, FL, USA) and spotted in duplicate in three drops of 333 pL each on 24-pad nitrocellulose-coated slides (ONCYTE AVID, GraceBio Labs, Bend, OR, USA) by using a non-contact Marathon Arrayjet microarray spotter (Roslin, UK).

Techniques: Recombinant, Microarray, Virus, Purification, SDS Page

Journal: Viruses

Article Title: Multiplex Serology for Sensitive and Specific Flavivirus IgG Detection: Addition of Envelope Protein Domain III to NS1 Increases Sensitivity for Tick-Borne Encephalitis Virus IgG Detection

doi: 10.3390/v16020286

Figure Lengend Snippet: Overview of serum collection used for orthoflavivirus microarray validation.

Article Snippet: TBEV rEDIII and the rNS1 of DENV 1 , DENV 2 , DENV 3 , DENV 4 , JEV, TBEV, USUV, WNV lineage 2 and ZIKV at concentrations of 1.5 mg/mL were mixed with 2× protein printing buffer (GVS, Sandford, FL, USA) and spotted in duplicate in three drops of 333 pL each on 24-pad nitrocellulose-coated slides (ONCYTE AVID, GraceBio Labs, Bend, OR, USA) by using a non-contact Marathon Arrayjet microarray spotter (Roslin, UK).

Techniques: Microarray, Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Virus, Neutralization

Journal: Viruses

Article Title: Multiplex Serology for Sensitive and Specific Flavivirus IgG Detection: Addition of Envelope Protein Domain III to NS1 Increases Sensitivity for Tick-Borne Encephalitis Virus IgG Detection

doi: 10.3390/v16020286

Figure Lengend Snippet: Validation of orthoflavivirus recombinant NS1 for sensitive and specific microarray detection of orthoflavivirus IgG. Heatmaps displaying microarray titers for the orthoflavivirus rNS1 of ( a ) orthoflavivirus negative reference sera, ( b ) WNV patients, ( d ) ZIKV patients, ( f ) DENV patients and ( h ) TBEV patients. (The color key of the titers is indicated at the bottom of each heatmap). The microarray antigens are listed on the left of each heatmap. Violin plots showing microarray titers of ( c ) WNV patients, ( e ) ZIKV patients, ( g ) DENV patients and ( i ) TBEV natural infection-positive sera against the recombinant NS1 of TBEV, DENV1, DENV2, DENV3, DENV4, ZIKV, WNV, USUV and JEV. A non-parametric t -test was performed to assess the statistical differences of paired samples. Significance is noted as ***, p < 0.001; *, p < 0.5. ( b ) Receiving operating characterization (ROC) curve was created with GraphPad Prism software (version 9.1.0).

Article Snippet: TBEV rEDIII and the rNS1 of DENV 1 , DENV 2 , DENV 3 , DENV 4 , JEV, TBEV, USUV, WNV lineage 2 and ZIKV at concentrations of 1.5 mg/mL were mixed with 2× protein printing buffer (GVS, Sandford, FL, USA) and spotted in duplicate in three drops of 333 pL each on 24-pad nitrocellulose-coated slides (ONCYTE AVID, GraceBio Labs, Bend, OR, USA) by using a non-contact Marathon Arrayjet microarray spotter (Roslin, UK).

Techniques: Biomarker Discovery, Recombinant, Microarray, Infection, Software

Journal: Viruses

Article Title: Multiplex Serology for Sensitive and Specific Flavivirus IgG Detection: Addition of Envelope Protein Domain III to NS1 Increases Sensitivity for Tick-Borne Encephalitis Virus IgG Detection

doi: 10.3390/v16020286

Figure Lengend Snippet: ( a ) Heatmap displaying microarray titers for the orthoflavivirus rNS1 of orthoflavivirus TBEV-vaccinated patients. (The color key of the titers is indicated at the bottom of the heatmap). The microarray antigens are listed on the left of the heatmap. ( b ) Violin plot showing microarray titers of the pre-characterized TBEV, dengue virus (DENV), Zika virus (ZIKV), West Nile virus (WNV) sera and the negative orthoflavivirus sera against TBEV rEDIII. TBEV vaccinees were used as the positive control. ( c ) Heatmap displaying microarray titers for the orthoflavivirus rEDIII of orthoflavivirus TBEV-vaccinated patients. (The color key of the titers indicated at the bottom of the heatmap.) The microarray antigens are listed on the y-axis. ( d )Violin plot showing microarray titers of TBEV-vaccinated serum samples against the recombinant EDIII of TBEV, DENV1, DENV2, DENV3, DENV4, ZIKV, WNV, USUV and JEV. A non-parametric T-test was performed to assess the statistical differences of paired samples. Significance is noted as ****, p < 0.0001; **, p < 0.01; *, p < 0.5. The order of patient samples is the same for each heatmap.

Article Snippet: TBEV rEDIII and the rNS1 of DENV 1 , DENV 2 , DENV 3 , DENV 4 , JEV, TBEV, USUV, WNV lineage 2 and ZIKV at concentrations of 1.5 mg/mL were mixed with 2× protein printing buffer (GVS, Sandford, FL, USA) and spotted in duplicate in three drops of 333 pL each on 24-pad nitrocellulose-coated slides (ONCYTE AVID, GraceBio Labs, Bend, OR, USA) by using a non-contact Marathon Arrayjet microarray spotter (Roslin, UK).

Techniques: Microarray, Virus, Positive Control, Recombinant

Journal: Viruses

Article Title: Multiplex Serology for Sensitive and Specific Flavivirus IgG Detection: Addition of Envelope Protein Domain III to NS1 Increases Sensitivity for Tick-Borne Encephalitis Virus IgG Detection

doi: 10.3390/v16020286

Figure Lengend Snippet: TBEV-infected serum samples against recombinant EDIII. ( a ) Heatmap displaying microarray titers for the orthoflavivirus rEDIII of orthoflavivirus TBEV natural infection-positive sera. (The color key of the titers is indicated at the bottom of each heatmap). The microarray antigens are listed on the y-axis. ( b ) Violin plots showing the microarray titers of TBEV natural infection-positive sera against the recombinant EDIII of TBEV, DENV1, DENV2, DENV3, DENV4, ZIKV, WNV, USUV and JEV. A non-parametric T-test was performed to assess the statistical differences of paired samples. Significance is noted as ****, p < 0.0001; ***, p < 0.001; *, p < 0.5. ( c ) Receiving operating characterization (ROC) curve was created with GraphPad Prism software (version 9.1.0). The order of patient samples is the same as in .

Article Snippet: TBEV rEDIII and the rNS1 of DENV 1 , DENV 2 , DENV 3 , DENV 4 , JEV, TBEV, USUV, WNV lineage 2 and ZIKV at concentrations of 1.5 mg/mL were mixed with 2× protein printing buffer (GVS, Sandford, FL, USA) and spotted in duplicate in three drops of 333 pL each on 24-pad nitrocellulose-coated slides (ONCYTE AVID, GraceBio Labs, Bend, OR, USA) by using a non-contact Marathon Arrayjet microarray spotter (Roslin, UK).

Techniques: Infection, Recombinant, Microarray, Software

Journal: Viruses

Article Title: Multiplex Serology for Sensitive and Specific Flavivirus IgG Detection: Addition of Envelope Protein Domain III to NS1 Increases Sensitivity for Tick-Borne Encephalitis Virus IgG Detection

doi: 10.3390/v16020286

Figure Lengend Snippet: Evaluation of recombinant orthoflavivirus EDIII as an antigen. Heatmaps and violin plots showing the microarray titers of ZIKV-positive sera ( a , b ), DENV-positive sera ( c , d ) and WNV-positive sera ( e , f ) against the recombinant rEDIII of TBEV, DENV1, DENV2, DENV3, DENV4, ZIKV, WNV, USUV and JEV. The order of patient samples is the same as in .

Article Snippet: TBEV rEDIII and the rNS1 of DENV 1 , DENV 2 , DENV 3 , DENV 4 , JEV, TBEV, USUV, WNV lineage 2 and ZIKV at concentrations of 1.5 mg/mL were mixed with 2× protein printing buffer (GVS, Sandford, FL, USA) and spotted in duplicate in three drops of 333 pL each on 24-pad nitrocellulose-coated slides (ONCYTE AVID, GraceBio Labs, Bend, OR, USA) by using a non-contact Marathon Arrayjet microarray spotter (Roslin, UK).

Techniques: Recombinant, Microarray

Journal: Viruses

Article Title: Multiplex Serology for Sensitive and Specific Flavivirus IgG Detection: Addition of Envelope Protein Domain III to NS1 Increases Sensitivity for Tick-Borne Encephalitis Virus IgG Detection

doi: 10.3390/v16020286

Figure Lengend Snippet: All TBEV serology results of sera ( n = 5) that tested TBEV IgG positive for rNS1 and/or rEDIII in the orthoflavivirus microarray.

Article Snippet: TBEV rEDIII and the rNS1 of DENV 1 , DENV 2 , DENV 3 , DENV 4 , JEV, TBEV, USUV, WNV lineage 2 and ZIKV at concentrations of 1.5 mg/mL were mixed with 2× protein printing buffer (GVS, Sandford, FL, USA) and spotted in duplicate in three drops of 333 pL each on 24-pad nitrocellulose-coated slides (ONCYTE AVID, GraceBio Labs, Bend, OR, USA) by using a non-contact Marathon Arrayjet microarray spotter (Roslin, UK).

Techniques: Microarray, Enzyme-linked Immunosorbent Assay, Infection