Journal: medRxiv

Article Title: Altered IgG4 Antibody Response to Repeated mRNA versus Protein COVID Vaccines

doi: 10.1101/2024.01.17.24301374

Figure Lengend Snippet: Randomly selected serum samples from COVID vaccine recipients included participants from 2019nCoV-307 ( ClinicalTrials.gov : NCT05463068 ) who received three homologous doses of Moderna (mRNA-1273, blue, n = 10) or Pfizer (BNT162b2, teal, n = 10) mRNA vaccine followed by one dose of Novavax (NVX-CoV2373), and from 2019nCoV-301 ( ClinicalTrials.gov : NCT04611802 ) who received four homologous doses of Novavax (purple, n = 18). Serum was collected ≥6 months after the last dose for homologous three-dose samples (open circle) and ∼4 weeks after fourth dose samples (filled circle). For each vaccine type, the GMFR between the third and fourth doses is indicated in bolded text above the fourth-dose data points. (A) Serum concentrations of anti–ancestral (Wuhan) rS–specific total IgG, IgG1, and IgG4 were measured using quantitative ELISA and reported as GMT with 95% CI. One subject (Moderna) was excluded due to a failed logic check. (B) ADCP, ADCC, and ADCD were analyzed using previously described surrogate SARS-CoV-2 ancestral rS–specific antibody Fc functional multiplex assays assessing median fluorescence intensity of FcγRIIa, FcγRIIIa, and C1q binding, respectively. One subject (Novavax) was excluded due to missing data (x3 Dose + Novavax). SAS ® 9.4 software (SAS Institute Inc., Cary, NC), PROC SGPLOT was used to detect far-outliers based on the log-transformed assay values, defined as Far-outliers = Observations > Q3 + 3*IQR or < Q1 – 3*IQR; none were found or excluded. Abbreviations: ADCC, antibody-dependent cellular cytotoxicity; ADCD, antibody-dependent complement deposition; ADCP, antibody-dependent cellular phagocytosis; CI, confidence interval; C1q complement component 1q; ELISA, enzyme-linked immunosorbent assay; FcγRIIa, Fcγ Receptor IIa; FcγRIIIa, Fcγ Receptor IIIa; GMFR, geometric mean fold rise; GMT, geometric mean titer; IgG, Immunoglobulin G; MFI, median fluorescent intensity; rS, recombinant spike protein; 3x, three-dose.

Article Snippet: The reference standards for ELISA were as follows; SARS-CoV-2 Spike RBD human IgG monoclonal antibody (Acro Biosystems, Cat# RAS009-C02) for total IgG, anti-SARS-CoV-2 Spike RBD human IgG1 monoclonal antibody (Acro Biosystems, Cat# SPD-M265) for IgG1, anti-SARS-CoV-2 spike RBD human IgG2 monoclonal antibody (Acro Biosystems, Cat# SPD-M400a), anti-SARS-CoV-2 spike RBD human IgG3 monoclonal antibody (Acro Biosystems, Cat# SPD-M401a), and anti-SARS-CoV-2 Spike RBD human IgG4 monoclonal antibody (Acro Biosystems, Cat# SPD-M402a) for IgG4.

Techniques: Enzyme-linked Immunosorbent Assay, Functional Assay, Multiplex Assay, Fluorescence, Binding Assay, Software, Transformation Assay, Recombinant

Journal: Vaccines

Article Title: COVID-19 Vaccination in Pregnancy: Pilot Study of Plasma MicroRNAs Associated with Inflammatory Cytokines after COVID-19 mRNA Vaccination

doi: 10.3390/vaccines12060658

Figure Lengend Snippet: Illustrates the experimental design of our pilot study. Peripheral blood samples were collected from pregnant women who received mRNA COVID-19 vaccines, and these samples were used for microRNA analysis and ELISA. The microRNA analysis was performed to identify differentially expressed microRNAs post-vaccination, while ELISA was used to measure the concentrations of S1RBD IgG and neutralizing antibodies against wild-type and different variants, and various cytokines, including IL-6, IL-6R, TNF-α, IL-10, and IL-1β (created using BioRender.com , accessed on 9 June 2024).

Article Snippet: The different variants of neutralizing antibodies, including the BA.1, BA.2, BA.4, and BA.5 variants, were determined using the GenScript SARS-CoV-2 Surrogate Virus Neutralization Test Kit for wild-type and AcroBiosystem Neutralizing Antibody Titer Serologic Assay Kit.

Techniques: Vaccines, Enzyme-linked Immunosorbent Assay