Subjects:

This study protocol will be registered with ClinicalTrails.gov and approval by an
independent ethics committee. All subjects will be provided detailed written information
concerning the trial, and will be required to give written consent to participate in the
study. The investigators will collect blood samples from residents associated with over
300 nursing homes, extended care facilities and over-55 communities which have agreed to
participate in the study at baseline (when available), and at 3, 6, 9 and 12 month
timepoints post-vaccination with one of the three authorized SARS-CoV-2 vaccines. The
plasma samples will be analyzed by Southlake Diagnostics.

The study will include up to 10,000 male and female residents of nursing homes, extended
care facilities and over-55 communities as well as staff associated with these
facilities. Demographic data of the subjects including characteristics, and medical
histories including concomitant medications and disease states/conditions will be
collected.

Blood Sample Collection:

A standard operating procedure (SOP) has been developed by Southlake Diagnostics to
establish a standardized procedure for establishing a safe and appropriate location and
method for obtaining human blood samples. This SOP incorporates regulations and
recommendations from the Occupational Safety and Health Administration (OSHA) Needlestick
Safety and Prevention Act (29 CFR 1910.1030). Blood draws conducted by Southlake
Diagnostics personnel will abide by this SOP.

The OSHA Bloodborne Pathogen Standard, (29 CFR 1910.1030) defines the required usage of
Safety Engineered Sharps Devices for any clinical research (including blood draws and
injections). All personnel conducting human blood draws are required to use Safety
Engineered Sharps Devices.

Personnel conducting blood draws are required to wear the appropriate personal protective
equipment (PPE). The Clinical Operations team is responsible for verifying that
laboratory personnel performing blood draws have appropriate training and experience in
conducting human blood sampling.

Blood samples will be drawn from an appropriate vein as the median cubital vein. Blood
samples of 3.0 ml will be collected in BD Vacutainer blood collection tubes containing
lithium-heparin using 21-23 gage needles. The blood samples will be packed in biohazard,
plastic leak-proof bags, and transported in insulated containers with cold packs to
ensure the integrity of the samples, as necessary. Plasma will be prepared by
centrifugation. Plasma samples will be stored at 2-8°C if not tested within 8 hours, and
frozen if not tested within 14 days.

Analytical Methods:

IgG antibodies to SARS-CoV-2 in human plasma (lithium-heparin) will be determined using
the Siemen's Atellica® IM SARS-CoV-2 IgG (sCOVG) assay (EUA Number: EUA202669) which is a
chemiluminescent immunoassay intended for qualitative and semi-quantitative detection of
IgG antibodies using the Atellica® IM Analyzer [1]. This SARS-CoV-2 IgG assay
specifically detects IgG antibodies to the S1 receptor binding domain (RBD) antigen. IgG
antibodies to SARS-CoV-2 are generally detectable in blood several days after initial
infection or vaccination. The duration of time antibodies is present post-vaccination is
not well characterized. The test will give an indication of the level of immunity that is
provided by a vaccine for the SARS-CoV-2 virus which causes COVID-19.

The Atellica IM sCOVG assay is a fully automated 2 step sandwich immunoassay using
indirect chemiluminescent technology. A direct relationship exists between the amount of
SARS-CoV-2 IgG antibody present in a patient plasma sample and the amount of relative
chemiluminescent light units (RLUs) detected by the system. Preparing master curves and
performing calibrations of the system will be performed according to manufacturing
instructions.

Results of the IgG antibody test are expressed as index values with an analytical
measuring interval of 0.50-150.00 Index units. Plasma samples with index values >150 will
be diluted and retested. Index values <1.00 will be considered negative (non-reactive)
while values >1.00 will be considered positive (reactive) [1].

Total (IgM + IgG) antibodies to SARS-CoV-2 in human plasma (lithium-heparin) will be
determined using the Siemen's Atellica® IM SARS-CoV-2 Total (COV2T) assay which is a
chemiluminescent immunoassay intended for qualitative detection of total antibodies using
the using the Atellica® IM Analyzer[1].

The Atellica IM COV2T assay is an automated immunoassay using indirect chemiluminescent
technology [1]. Results of the total (IgM + IgG) antibody test are expressed as index
values with an analytical measuring interval of 0.05-10.00 Index units. Index values
<1.00 will be considered negative (non-reactive) while values >1.00 will be considered
positive (reactive) [1]

[1].
https://www.siemens-healthineers.com/en-us/laboratory-diagnostics/assay…
itions/infectious-disease-assays/sars-cov-2-igg-assay

Data Management:

Demographic data of the subjects including characteristics, and medical histories
including concomitant medications and disease states/conditions will be provided for each
subject by the respective residence and inputted into the REDCap cloud platform
(described below) by data input specialists employed by Southlake Diagnostics. Data entry
will be checked by a second specialist for accuracy of entry. Standardized and approved
terminology will be used for all entry parameters as height, weight, age, drug names, and
disease states to ensure consistency of data entry. Data entry will be reviewed for
accuracy of entry and validation by the Co-Principal Investigator (Dr. Robert Newton) who
is responsible for this function, and the Principal Investigator.

Statistical Methods:

The laboratory will provide the certified statistician with a spread sheet of the data
which will be sorted and analyzed as a whole and according to various parameters as
gender, age groups, known disease states and immunocompetencies, physical characteristics
and time after vaccination using a REDCap Cloud platform (www.redcapcloud.com/). This
platform is advanced, compliant software for the management of all types of data
associated with clinical studies. The system is Medical Dictionary for Regulatory
Activities (MEdDRA) terminology certified, Clinical Data Interchange Standards Consortium
(CDISC) compliant, and International Standards Organization (ISO) 27001 certified. The
data will be sorted and analyzed for statistical significance and trends using various
computer programs associated with the platform.

Information regarding overall duration and rate of decline of antibody levels as a
function of time post-vaccination as well as the effects of confounding factors on
antibody levels as a function of time will be determined. Information regarding antibody
levels (IgG and total) and Covid-19 infection or reinfection, and approximate time at
which a booster shot may be required will also be obtained relative to antibody levels.
The large number of subjects in the overall study should enable statistical associations
to be made between antibody levels and various potentially confounding and influencing
factors. The data will be reviewed by the Principal Investigator and Co-PIs.