Alignment of the amino acid sequences containing the fusion loop domain from E proteins of DENV 1–4, WNV, JEV, YFV, TBEV and the four point mutations of the QUAD proteins.

<p>Amino acids numbering: 1 is start of the E protein.</p

IgM-ELISA on 300 ng of DENV-2 Ewt (A) and DENV 1–4 Equad mixture (B) with different DENV, WNV and negative (NEG) sera (n = number of individuals).

<p>Bottom and top of the boxes are the first and third quartiles. The median signal is depicted as a line inside the box. Whiskers represent the 9^th and the 91^st percentile. Measurements were performed in duplicates in at least two independent experiments.</p

Comparison of different antigens for the detection of DENV IgG.

<p>Sera positive for IgG against DENV, WNV, TBEV or negative control sera were analyzed with the Panbio Indirect IgG ELISA (black), the DENV-2 Ewt protein (light gray, lined) or the DENV1-4 Equad mix (dark grey, lined). The absolute absorbance is indicated. Cut-Off values for the Panbio test were obtained by calculation of the internal standard of the manufacturer; these are indicated at the right and only refer to this test (horizontal bars: DENV-positive results with an OD-value higher than 1.1*cut-off, equivocal results having an OD-value between 1.1*cut-off and 0.9*cut-off, negative results with an OD-value lower than 0.9*cut-off).</p

Antigen titration using the indicated amounts of DENV-1 (A), -2 (B), -3 (C) and -4 (D) Equad and DENV-2 Ewt (E) proteins with three different DENV, WNV and negative (NEG) human sera.

<p>Measurements were performed in duplicates in at least two independent experiments.</p

A: Expression and purification of DENV-2 Ewt and Equad from Drosophila S2 culture supernatants; supernatant before induction (b.ind.), after 7 days of expression culture (7 d expr.), concentrated via tangential flow (Conc.) and the two step purification with immobilized imidazole affinity (IMAC) and size exclusion chromatography (SEC) were separated on a 10% SDS-PAGE gel under reducing conditions. B: 6 μg of purified DENV1-4 (D1-D4) Equad and DENV-2 Ewt proteins were analyzed with SDS-PAGE. Proteins were stained with Coomassie blue. Size of molecular weight markers in kilo Daltons is indicated on the left.

<p>A: Expression and purification of DENV-2 Ewt and Equad from Drosophila S2 culture supernatants; supernatant before induction (b.ind.), after 7 days of expression culture (7 d expr.), concentrated via tangential flow (Conc.) and the two step purification with immobilized imidazole affinity (IMAC) and size exclusion chromatography (SEC) were separated on a 10% SDS-PAGE gel under reducing conditions. B: 6 μg of purified DENV1-4 (D1-D4) Equad and DENV-2 Ewt proteins were analyzed with SDS-PAGE. Proteins were stained with Coomassie blue. Size of molecular weight markers in kilo Daltons is indicated on the left.</p

300 ng per well of DENV-2 Ewt (A) and Equad (B) and 160 ng per well of a DENV 1–4 Equad mixture (C) were tested with DENV- WNV- and TBEV- infected and YFV-vaccinated sera compared to negative (NEG) samples in an IgG-ELISA (n = number of individuals).

<p>Bottom and top of the boxes are the first and third quartiles. The median signal is depicted as a line inside the box. Whiskers represent the 9^th and the 91^st percentile. Outliers in B and C are marked with numbers (1–5). Measurements were performed in duplicates in at least two independent experiments.</p

BNT162b2- and infection induced reciprocal SARS-CoV-2 nAb titers on wildtype strain wt-BavPat1 and VOC strains Delta, Beta and Omicron at timepoint P1 to timepoint P3.

The dotted line indicates the limit-of-detection at a titer of 1:5. FRNT90: Focus reduction neutralization titer at 90% virus inhibition; results plotted as reciprocal values. Proportion of individuals with remaining nAbs in percent. Mean neutralizing titer reductions of SARS-CoV-2 wt to VOC-nAb are depicted above the continuous lines * = pppp<0.0001, ns = not significant.</p

Salivary IgA specific to SARS-CoV-2 RBD in N = 40 individuals prior first (V1), three weeks after first (V2) and to weeks after second vaccination (P1) in comparison with COVID-19 patients (N = 31).

The dotted line indicates the 95% interquartile range for vaccinees.</p

Neutralizing antibodies in ChAdOx1-S/BNT162b2 vaccinated individuals to SARS-CoV-2 wild-type and VOCs.

(PDF)</p

Time points of blood sampling within the different cohorts in median days Vaccinees: V1 prior first vaccination, V2 21 days after first vaccination, P1 21 days, P2 60 days and P3 240 days after second vaccination.

Boost samples were taken 14 days after third vaccination. BNT162b2 vaccinees received their second vaccination 3 weeks after the first dose, whereas heterologous vaccinees received their second dose (BNT16b2) 10 weeks after the first vaccination with ChAdOx1-S. COVID-19 Patients: 35 days post symptom onset (PSO).</p