Set of data investigating the immunogenicity of SARS-CoV-2 spike protein.

GraphPad Prism, 8.0.2

METHODOLOGICAL INFORMATION

1. Description of methods used for collection-generation of data: Humoral Response Section: Blood samples were collected from mice on days 21, 35, and 56 post-initial injection to evaluate their humoral response to the protein antigen. Antibody levels were assessed using the Recombivirus Mouse Anti SARS-CoV-2 (COVID) Spike 1 RBD IgG ELISA Kit (RV-405420, Alpha Diagnostics, San Antonio, Texas, U.S.A.) following the manufacturer's instructions. In Vitro Neutralization Assay Section:For the in vitro neutralization assay, 7.5 μL of each animal's serum was incubated with 20 or 50 plaque-forming units of SARS-CoV2 (Strain: hCoV19/Spain/SP-VHIR.02, D614G(S)) in a biosafety level 3 environment. Each serum sample was tested in five independent replicates. After a one-hour incubation, the pre-incubated serum was added to VeroE6 cell-containing 24-well plates. Following a six-hour incubation period, the cells were washed to remove any residual SARS-CoV-2 particles. Subsequently, at either 24 or 48 hours post-infection, aliquots were taken, centrifuged, and the supernatant was inactivated with RNA Shield. Viral RNA extraction was then performed, followed by reverse transcription and RT-qPCR analysis using specific primers. The results, expressed as Cycle Quantification (Cq) values, indicate the initial viral load, with lower values reflecting higher viral RNA concentrations. Procedure HR632-21 was approved by the Biosafety Committee of the UAB. The rest of methods are standard and well described elsewhere: https://doi.org/10.3390/pharmaceutics15041197 and https://doi.org/10.1021%2Facsami.3c08643

2. Methods for processing the data: Data processing details are standard and well described elsewhere: https://doi.org/10.3390/pharmaceutics15041197

3. Instrument- or software- specific information needed to interpret the data: GraphPad Prism, version 8.0.2, available at: https://www.graphpad.com/

4. Instruments, calibration and standards information: EmulsiFlex-C5 system (Avestin) ÄKTA pure system (Cytiva) Zetasizer Pro (Malvern Analytical)

5. Environmental or experimental conditions: Data processing and figure creation were performed in Windows 10 OS.