Journal: Frontiers in Immunology

Article Title: Humoral Immune Response Diversity to Different COVID-19 Vaccines: Implications for the “Green Pass” Policy

doi: 10.3389/fimmu.2022.833085

Figure Lengend Snippet: Spearman’s correlation analysis of serum anti-receptor-binding domain (anti-RBD) antibody titers and neutralizing activity in 70 participants in the study (22 vaccinated with BNT162b2, 9 with mRNA-1273, 27 with ChAdOx1 nCov19, 9 with Ad26.COV2.S, 1 COVID-19 convalescent, and 2 with mixed vaccines). (A) Correlation plot of anti-RBD antibody titers versus neutralizing activity (percentage inhibition of RBD-ACE2 binding) assessed through the cPass™ ELISA-based assay. (B) Correlation plot of anti-RBD antibody titers versus neutralizing activity assessed through IgG/Neutralizing Antibody Rapid Test. (C) Correlation plot of neutralizing activity evaluated through cPass™ ELISA-based assay and IgG/Neutralizing Antibody Rapid Test cassettes. Trendlines, Spearman’s r , and p -values are also represented (statistical significance for p < 0.05).

Article Snippet: Some of them have high costs, require trained personnel, and can only be carried out in a Biosafety Safety Level 3-equipped laboratory, whereas others, such as the cPass surrogate virus neutralization test (sVNT) (GenScript, Piscataway, NJ, USA) used in this work, are ELISA-based assays and only require optical density readers.

Techniques: Binding Assay, Activity Assay, Vaccines, Inhibition, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Humoral Immune Response Diversity to Different COVID-19 Vaccines: Implications for the “Green Pass” Policy

doi: 10.3389/fimmu.2022.833085

Figure Lengend Snippet: Neutralizing activity evaluated by cPass™ ELISA-based SARS-CoV-2 Neutralization Antibody Detection Kit in 70 sera from differently vaccinated individuals. Serum samples were considered positive when ≥30% inhibition was measured, as shown by the red line in the graph. (A) Percentage inhibition of receptor-binding domain–angiotensin-converting enzyme 2 (RBD-ACE2) binding within different vaccination groups (see also Table 3 ). Statistical significance was assessed by ANOVA following Tukey’s multiple comparisons test, ** p < 0.005, * p < 0.05. (B) Comparison of neutralizing activity in sera from individuals who received adenoviral DNA-based vaccines and mRNA-based vaccines. Statistical significance was assessed by unpaired t-test, * p = 0.0016.

Article Snippet: Some of them have high costs, require trained personnel, and can only be carried out in a Biosafety Safety Level 3-equipped laboratory, whereas others, such as the cPass surrogate virus neutralization test (sVNT) (GenScript, Piscataway, NJ, USA) used in this work, are ELISA-based assays and only require optical density readers.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Neutralization, Inhibition, Binding Assay, Comparison, Vaccines

Journal: Frontiers in Immunology

Article Title: Humoral Immune Response Diversity to Different COVID-19 Vaccines: Implications for the “Green Pass” Policy

doi: 10.3389/fimmu.2022.833085

Figure Lengend Snippet: Mean neutralizing activity measured by the cPass™ ELISA-based assay in sera from 70 individuals who received different vaccines.

Article Snippet: Some of them have high costs, require trained personnel, and can only be carried out in a Biosafety Safety Level 3-equipped laboratory, whereas others, such as the cPass surrogate virus neutralization test (sVNT) (GenScript, Piscataway, NJ, USA) used in this work, are ELISA-based assays and only require optical density readers.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Vaccines

Journal: medRxiv

Article Title: Long-term immunogenicity of BNT162b2 vaccination in the elderly and in younger health care workers

doi: 10.1101/2021.08.26.21262468

Figure Lengend Snippet: (A) Anti-S1 IgG in serum measured by a microarray-based immunoassay, (B) serum pseudovirus neutralization against the Delta VOC two and six months after vaccination measured by pNT, and (C) SARS-CoV-2 S1 specific T cell response detected by IGRA. Dotted horizontal lines indicate themanufacturer’s threshold for anti-S1 IgG ≥1 S/Co (A) and the lower limit of detection (1:10 dilution) for pNT (B). Horizontal lines within plotted data regions indicate the median and interquartile range, except for pNT, where the geometric mean and 95% confidence interval is shown. P values (all <0.0001) were calculated by non-parametric Mann Whitney U test. S/Co: signal-to-cutoff, pNT: pseudovirus neutralization test, ID 50 : 50% inhibition dilution, IGRA: interferon-γ release assay, IU: international units.

Article Snippet: Functional neutralization capacity was investigated using a commercially available ELISA-based SARS-CoV-2 RBD-ACE2 binding inhibition assay (surrogate SARS-CoV-2 neutralization test (sVNT) cPass (medac GmbH, https://international.medac.de ).

Techniques: Microarray, Neutralization, MANN-WHITNEY, Inhibition, Release Assay

Journal: medRxiv

Article Title: Long-term immunogenicity of BNT162b2 vaccination in the elderly and in younger health care workers

doi: 10.1101/2021.08.26.21262468

Figure Lengend Snippet: ( A ) Anti-SARS-CoV-2 N, ( B ) RBD- ( C ) and full-spike IgG measured in the serum of BNT162b2 vaccinated HCW and elderly persons six months after the first vaccination. ( D ) Neutralizing capacity was measured by sVNT and ( E ) serum neutralization against Alpha (B.1.1.7) VOC detected by pNT in vaccinated HCW and elderly persons six months after the first vaccination. ( F ) Binding capacity of serum IgG against six different RBDs of SARS-CoV-2 variants carrying the indicated mutations in HCW and elderly, measured by ELISA. Dotted lines indicate the manufacturer’s threshold values: for anti-N, anti-RBD, and anti-full spike IgG ≥1 S/Co, for sVNT >30%, and the lower limit of detection (1:10 dilution) for pNT. Lines indicate the median and interquartile range except for pNT, where the geometric mean and 95% confidence interval are shown. P values were calculated by the non-parametric Mann Whitney U test or Kruskal-Wallis test with Dunn’s multiple comparisons test. S/Co: signal-to-cutoff, N: nucleocapsid protein, RBD: receptor-binding domain, sVNT: surrogate virus neutralization test, ACE2: angiotensin-converting enzyme 2, ID50: 50% inhibition dilution.

Article Snippet: Functional neutralization capacity was investigated using a commercially available ELISA-based SARS-CoV-2 RBD-ACE2 binding inhibition assay (surrogate SARS-CoV-2 neutralization test (sVNT) cPass (medac GmbH, https://international.medac.de ).

Techniques: Neutralization, Binding Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Inhibition

Journal: Diagnostics

Article Title: Anti-SARS-CoV-2 IgM Antibody Levels Measured by an In-House ELISA in a Convalescent Latin Population Persist over Time and Exhibit Neutralizing Capacity to Several Variants of Concern

doi: 10.3390/diagnostics14192209

Figure Lengend Snippet: Agreement between the results obtained by the in-house CovIgM-ELISA and the surrogate neutralization assay (cPass).

Article Snippet: Having previously demonstrated that the results provided by the surrogate viral neutralization test (cPass TM GenScript sVNT, Piscataway NJ, USA) [ ] correlate perfectly with the traditional PRNT [ ], a cPass neutralization antibody test was used to determine the levels of neutralizing antibodies in the study population against the wild-type strain and the variants of concern (VOCs), Alpha, Delta and Omicron, which had a wide circulation and prevalence in the Puerto Rican population [ ]. cPass utilizes the recombinant RBD of the SARS-CoV-2 spike protein to detect antibodies that block the RBD from binding to the human ACE2 receptor.

Techniques: Neutralization

Journal: Diagnostics

Article Title: Anti-SARS-CoV-2 IgM Antibody Levels Measured by an In-House ELISA in a Convalescent Latin Population Persist over Time and Exhibit Neutralizing Capacity to Several Variants of Concern

doi: 10.3390/diagnostics14192209

Figure Lengend Snippet: Pearson (r) correlation between the neutralization percentages (sVNT%) and levels of anti-SARS-CoV-2 IgM . The levels of anti-SARS-CoV-2 IgM expressed as average OD 450nm for each sample determined by the in-house CovIgM-ELISA was associated with the neutralization assay (cPass, Genscript) against the wild-type strain and the variants of concern Alpha, Delta and Omicron. Dashed lines on x-axis represent the positive cut-off value (OD > 0.26) for in-house CovIgM-ELISA and the dashed line on the y-axis represent the positive cut-off (sVNT% ≥ 30%) for the neutralization assay. There was significant correlation (* p = 0.02225) between sVNT% and anti-SARS-CoV-2 IgM levels for the wild-type strain.

Article Snippet: Having previously demonstrated that the results provided by the surrogate viral neutralization test (cPass TM GenScript sVNT, Piscataway NJ, USA) [ ] correlate perfectly with the traditional PRNT [ ], a cPass neutralization antibody test was used to determine the levels of neutralizing antibodies in the study population against the wild-type strain and the variants of concern (VOCs), Alpha, Delta and Omicron, which had a wide circulation and prevalence in the Puerto Rican population [ ]. cPass utilizes the recombinant RBD of the SARS-CoV-2 spike protein to detect antibodies that block the RBD from binding to the human ACE2 receptor.

Techniques: Neutralization, Enzyme-linked Immunosorbent Assay

Journal: Vaccine

Article Title: Semi-quantitative, high throughput analysis of SARS-CoV-2 neutralizing antibodies: Measuring the level and duration of immune response antibodies post infection/vaccination

doi: 10.1016/j.vaccine.2021.07.098

Figure Lengend Snippet: Longitudinal analysis of neutralizing antibodies pre and post-vaccination with cPass sVNT. A. Qualitative (% Neutralization) assessment of neutralizing antibodies with Pfizer and Moderna vaccines. Samples were tested before (pre-vaccination), or within two to three weeks after each dose (post-1st or 2nd dose). B. Qualitative (% Neutralization) analysis of neutralizing antibodies with Johnson & Johnson vaccine. Samples were tested before (pre-vaccination), two weeks after and 39 days after vaccination. C. Semi-quantitative testing of neutralizing antibody titers post-vaccination. Samples were collected from the same individuals over multiple time points post-vaccination and assessed semi-quantitatively. D. Direct comparison between qualitative (% neutralization) and semi-quantitative (ng/ml titers) post-vaccination. Samples were collected from subject 152 ( C) using the cPass sVNT qualitative and semi-quantitative protocols with semi-quantitative data plotted on the left Y-axis and qualitative data on the right Y-axis.

Article Snippet: This study describes an approach for accurate quantification of neutralizing antibodies using the cPass sVNT with an automated workflow on the Tecan EVO and Dynex Agility platforms that is applicable to other liquid handling systems.

Techniques: Neutralization, Vaccines, Comparison

Journal: Vaccine

Article Title: Semi-quantitative, high throughput analysis of SARS-CoV-2 neutralizing antibodies: Measuring the level and duration of immune response antibodies post infection/vaccination

doi: 10.1016/j.vaccine.2021.07.098

Figure Lengend Snippet: Optimization of cPass sVNT for liquid handler automation. A. Ambient temperature incubations. Ten replicates of a standard curve using the kit insert (KI) “Standard Protocol” were compared to nine replicates from an optimized ambient temperature “Room Temp” procedure (see section 2.5) using a well-characterized neutralizing monoclonal antibody (MAB) (GenScript #A02051) blocking the RBD-ACE2 interaction. The MAB was diluted serially by a factor of 1:2 from 150 ng/mL down each column of the plates to assess and compare intra-plate variability. The data between the two procedures were compared by plotting both the raw (OD450) values (A1 and A3) and the transformed % neutralization results (A2 and A4) vs Log of Ab Concentration. Within the linear range (ie: the second and third MAB dilutions) the difference between the standard (manual) and Tecan (automated) protocols was within 20% CV. B. Effect and compensation of wash buffer dilution of TMB. Mixtures of 20 µL residual wash buffer with the KI recommended 100 µL as well as 150 µL and 200 µL TMB were compared with the KI protocol of no residual wash buffer with 100 µL TMB. The data between the two procedures were compared by plotting both the raw (OD450) values (B2) and the transformed % neutralization results (B1) vs Log of Ab Concentration. Using a 2-way, multiple comparisons ANOVA test, a statistically significant difference was revealed within the linear range between the baseline (100 µL TMB) and the other conditions that included wash buffer with the exception of “20 µL wash buffer + 200 µL TMB” where there was no statistically significant difference.

Article Snippet: This study describes an approach for accurate quantification of neutralizing antibodies using the cPass sVNT with an automated workflow on the Tecan EVO and Dynex Agility platforms that is applicable to other liquid handling systems.

Techniques: Blocking Assay, Transformation Assay, Neutralization, Concentration Assay

Journal: Vaccine

Article Title: Semi-quantitative, high throughput analysis of SARS-CoV-2 neutralizing antibodies: Measuring the level and duration of immune response antibodies post infection/vaccination

doi: 10.1016/j.vaccine.2021.07.098

Figure Lengend Snippet: Qualitative analysis of 86 SARS-CoV-2 positive and pre-pandemic samples using cPass sVNT, Platelia SARS-CoV-2 Total Ab and Lumit™ Dx SARS-CoV-2 Immunoassay tests. The identical set of 57 positive and 29 pre-pandemic samples were screened qualitatively using cPass sVNT, Platelia SARS-CoV-2 Total Ab and Lumit™ Dx SARS-CoV-2 Immunoassay tests. The same two positive samples were delineated negative (ie: false negative) for the cPass and Lumit assays and one of the false negative samples were in common with all three kits (red dots). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: This study describes an approach for accurate quantification of neutralizing antibodies using the cPass sVNT with an automated workflow on the Tecan EVO and Dynex Agility platforms that is applicable to other liquid handling systems.

Techniques:

Journal: Vaccine

Article Title: Semi-quantitative, high throughput analysis of SARS-CoV-2 neutralizing antibodies: Measuring the level and duration of immune response antibodies post infection/vaccination

doi: 10.1016/j.vaccine.2021.07.098

Figure Lengend Snippet: Combined data from the qualitative population surveillance study ( Fig. 2 ) summarizing assay performance characteristics of cPass sVNT, Platelia SARS-CoV-2 Total Ab and Lumit™ Dx SARS-CoV-2 Immunoassay tests. Sensitivity, specificity, accuracy, positive and negative predictive values are shown. A prevalence of 10% was used for the calculations.

Article Snippet: This study describes an approach for accurate quantification of neutralizing antibodies using the cPass sVNT with an automated workflow on the Tecan EVO and Dynex Agility platforms that is applicable to other liquid handling systems.

Techniques: