Communication

A Peptide-Based Assay Discriminates Individual Antibody

Response to the COVID-19 Pfizer/BioNTech mRNA Vaccine
Immacolata Polvere 1,2 , Serena Voccola 2,3 , Alfredina Parrella 3 , Gaetano Cardinale 3 , Lucrezia Zerillo 1,2 ,
Romualdo Varricchio 2 , Jessica Raffaella Madera 1 , Romania Stilo 1 , Pasquale Vito 1,2, * and Tiziana Zotti 1,2, *

1 Dipartimento di Scienze e Tecnologie, Università Degli Studi del Sannio, Via dei Mulini,
82100 Benevento, Italy; immapolvere88@gmail.com (I.P.); lzerillo@unisannio.it (L.Z.);
jrmadera@unisannio.it (J.R.M.); romsti@unisannio.it (R.S.)
2 Genus Biotech, Università Degli Studi del Sannio, SS Appia, 82030 Apollosa, Italy;
serena.voccola@tecnobios.com (S.V.); romualdovar@outlook.it (R.V.)
3 Consorzio Sannio Tech, SS Appia, 82030 Apollosa, Italy; alfredina.parrella@tecnobios.com (A.P.);
gaetano.cardinale@tecnobios.com (G.C.)
* Correspondence: vito@unisannio.it (P.V.); tzotti@unisannio.it (T.Z.)






Citation: Polvere, I.; Voccola, S.;
Parrella, A.; Cardinale, G.; Zerillo, L.;
Varricchio, R.; Madera, J.R.; Stilo, R.;
Vito, P.; Zotti, T. A Peptide-Based
Assay Discriminates Individual
Antibody Response to the COVID-19
Pfizer/BioNTech mRNA Vaccine.
Vaccines 2021, 9, 987. https://
doi.org/10.3390/vaccines9090987
Academic Editor: Siddappa
N. Byrareddy
Received: 21 July 2021
Accepted: 2 September 2021
Published: 3 September 2021
Abstract: The coronavirus disease 2019 (COVID-19) mRNA vaccine developed by Pfizer/BioNTech
has been shown to be capable of developing an excellent antibody response against the severe
acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein, with good production of
neutralizing antibodies. Herein, we analyzed differences in the antibody response elicited by inocula-
tion of the Pfizer/BioNTech vaccine through a peptide-based enzyme-linked immunosorbent assay
(ELISA) that utilizes synthetic peptides derived from the spike protein in the immuno-adsorbent
phase. Immunoreactivity against synthetic peptides was measured at different time points from
vaccination and was also correlated with the SARS-CoV-2 neutralizing capacity. Our results indicate
that all vaccinated subjects except one show reactive antibodies to at least one peptide at both 30 and
60 days after injection of the first dose. Only one of the 19 analyzed subjects showed no antibody
response toward any of the selected peptides, consistently with a lower neutralizing capacity. More
importantly, our data showed that the antibody response elicited by inoculation of the two doses of
the Pfizer vaccine appears to be qualitatively individual, both in the type of recognized peptides and
in the temporal persistence of the antibody response. Together with previous published data, our
findings suggest that for effective pandemic control, it is important to constantly monitor the antibody
protection in the population, and the assay described here could be a valid tool for this purpose.
Keywords: COVID-19; vaccine; antibodies; ELISA; peptides

1. Introduction
Publisher’s Note: MDPI stays neutral
More than 18 months after the outbreak of the COVID-19 pandemic due to the SARS-
with regard to jurisdictional claims in
CoV-2 infection, which has severely tested the health and economic structure of the world,
published maps and institutional affil-
there is a shared belief that the introduction of mass vaccination may actually lead to a
iations.
control of the spread of the virus [1,2]. From this perspective, the design, development,
validation, and administration to large populations of different vaccines against COVID-19
in an exceptionally short period of time can be considered one of the greatest achievements
of all time in medical history [3]. Among all COVID-19 vaccines, whether prepared or in
Copyright: © 2021 by the authors.
development, including attenuated, inactivated, subunit, and nucleic acid-based vaccines,
Licensee MDPI, Basel, Switzerland.
those based on mRNA molecules have aroused great interest for their novelty and for the
This article is an open access article
exceptional efficacy observed in the clinical trial phase [2,3].
distributed under the terms and
BNT162b2 developed by Pfizer/BioNTech is a COVID-19 vaccine containing nucleoside-
conditions of the Creative Commons
modified messenger RNA encoding the SARS-CoV-2 spike glycoprotein [4]. In healthy
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
adults and young adults older than 16, two 30 µg doses of BNT162b2 elicit high neutraliz-
4.0/).
ing titers and robust, antigen-specific T cell responses against SARS-CoV-2, and are 95%

Vaccines 2021, 9, 987. https://doi.org/10.3390/vaccines9090987 https://www.mdpi.com/journal/vaccines

Vaccines 2021, 9, 987 2 of 7

effective in preventing severe disease due to virus infection from seven days after the
second dose [5,6]. In adolescents of 12–15 years of age, the vaccine’s efficacy was 100% [7].
We previously reported the development of a peptide-based ELISA by selecting seven
synthetic peptides from the spike, membrane, and nucleocapsid protein sequences of
SARS-CoV-2, which effectively detects the antibody response to SARS-CoV-2 mounted
by COVID-19 convalescents [8]. The assay detects a significant difference in the antibody
response among single subjects, proving that the antibody responses to SARS-CoV-2 in
infected patients appears to have unique repertoire distribution patterns without pref-
erence for particular antibody families. Therefore, using the same peptide-based ELISA
assay, we verified whether the antibody response elicited by the Pfizer/BioNTech vaccine
inoculation was either heterogeneous, similarly to the one elicited by the viral infection, or,
alternatively, more homogeneous, being the vaccine based on the expression of the viral
spike protein alone.

2. Materials and Methods

2.1. Study Design and Informed Consent
All study participants (19 individuals: 12 females, 7 males; aged 25–70 years, median
32 years), who also include the authors of this manuscript, are part of the staff of a
diagnostic center for COVID-19. Participants received two doses of the Pfizer/BioNTech
vaccine (first dose at day 0 and second dose at day 21) and voluntarily underwent venous
blood sampling for serological tests at 0, 15, 30, and 60 days after the first dose, in order to
monitor the presence of anti-SARS-CoV-2 neutralizing antibodies. All participants signed
an informed consent for the anonymized use of the leftover blood sample. None of the
participants, except vax_8, had a previous history of COVID-19. The study was approved
by the Institutional Review Board of Consorzio Sannio Tech (n. 01/2021).

2.2. Blood Samples

After clotting, serum was separated using centrifugation for 10 min at 1000 rcf and
heat-inactivated for 30 min at 56 ◦ C. Then, 500 µL aliquots were stored at –80 ◦ C.

2.3. ELISA Assay

The peptides used in this work are reported in Table 1 and ELISA was performed as
published elsewhere [8]. Briefly, coating of 96-well high-binding plates (NUNC Maxisorp,
Thermo Scientific, USA) with 2 µg/mL of single or pooled peptides in HBSS was performed
overnight at 4 ◦ C. Sera samples were diluted in blocking buffer (1:500) and incubated
for 1 h at room temperature with continuous agitation. Then, the plates were washed
and incubated with 1:60,000 diluted horseradish peroxidase (HRP)-conjugated goat anti-
human immunoglobulin G (MERCK KGaA, Darmstadt, Germany). After washing, color-
developing reaction was carried out for 15–30 min at room temperature in the dark by
adding 70 µL of 1:3 freshly diluted 1-StepTM Ultra TMB-ELISA (Thermo Scientific, Waltham,
MA, USA) and stopped with an equal volume of 0.3 M H2 SO4 . Absorbance readings at
450 nm were taken using the microplate reader Seac-Sirio-S. Two pre-2019 sera were used
as negative controls, while seropositive controls were obtained from two different patients
who had been infected with SARS-CoV-2 in the four months prior the study.

Table 1. List of peptides used for this study.

Peptide Sequence Position

Pep2_Spike DAVDCALDPLSETKCTLKSFTVEKGIYQTSN 287–317
Pep5_Spike FSQILPDPSKPSKRSFIE 802–819
Pep6_Spike GTNTSNQVAVLYQDVNCTEVPVAIHADQLTPTWRVYSTGS 601–640
VCGPKKSTNLVKNKCVNFNFNGLTGTGVLTESNKKFLPFQQFGRDI
Pep10_Spike 524–598
ADTTDAVRDPQTLEILDITPCSFGGVSVI
Vaccines 2021, 9, 987 3 of 7

Qualitative direct detection of total neutralizing antibodies to SARS-CoV-2 in human

serum was performed with a cPassTM SARS-CoV-2 Neutralization Antibody Detection
Kit (GenScript Biotech Corporation, Piscataway Township, NJ, USA) according to the
manufacturer’s instructions.

3. Results and Discussion

Four peptides (Table 1) derived from the spike protein of the SARS-CoV-2 Hu-1 strain
(GeneBank: MN908947) were used as the immuno-adsorbent phase in an ELISA. These
peptides are generally seen as targets by the antibody response generated following virus
infection [8].
Figure 1 shows the IgG antibody response mounted by 19 individuals receiving
the first dose of the Pfizer/BioNTech vaccine at day 0 and the second dose at day 21.
The humoral response was monitored at days 15, 30, and 60 from day 0. Instead of the
signal/cutoff ratio used as a scoring method in previous studies [8], the antibody response
is reported as a log2 fold-change in the ELISA absorbance readout compared to day 0,
which is more useful when analyzing variations through multiple measurements of a
Vaccines 2021, 9, 987 4 of 8
biological parameter in the same individual at different time points. In order to better
discriminate differences in humoral responses over time, positivity was arbitrarily scored
for a log2 fold-change >1.5.

15days
30days
Reactivity to pep2-Spike Reactivity to pep5-Spike 60days
16 16

8
8
Fold Change (log2)
Fold Change (log2)

4
4
2

2
1

1 0.5

0.5 0.25
vax_1
vax_2
vax_3
vax_4
vax_5
vax_6
vax_7
vax_8
vax_9
vax_10
vax_11
vax_12
vax_13
vax_14
vax_15
vax_16
vax_17
vax_18
vax_19
vax_1
vax_2
vax_3
vax_4
vax_5
vax_6
vax_7
vax_8
vax_9
vax_10
vax_11
vax_12
vax_13
vax_14
vax_15
vax_16
vax_17
vax_18
vax_19

Reactivity to pep6-Spike Reactivity to pep10-Spike

32 4

16
Fold Change (log2)

Fold Change (log2)

8 2

2 1

0.5 0.5
vax_1
vax_2
vax_3

vax_4
vax_5
vax_6
vax_7
vax_8
vax_9
vax_10
vax_11
vax_12

vax_13
vax_14
vax_15
vax_16
vax_17
vax_18
vax_19

vax_1
vax_2
vax_3

vax_4
vax_5
vax_6
vax_7
vax_8
vax_9
vax_10
vax_11
vax_12

vax_13
vax_14
vax_15
vax_16
vax_17
vax_18
vax_19

(a)

Figure 1. Cont.
 Vaccines 2021, 9, 987 5 of 8
Vaccines 2021, 9, 987 4 of 7

Pep2_Spike Pep5_Spike Pep6_Spike Pep10_Spike

T15
T30
T60

T15
T30
T60

T15
T30
T60

T15
T30
T60
vax_1
vax_2
vax_3
vax_4
vax_5
vax_6
vax_7
vax_8
vax_9
vax_10
vax_11
vax_12
vax_13
vax_14
vax_15
vax_16
vax_17
vax_18
vax_19

(b)
Figure
Figure 1. IgG
1. (a) (a) IgG immunoreacting
immunoreacting with with the selected
the selected peptides
peptides at and
at 15, 30, 15, 30, and following
60 days 60 days following Pfizer/BioNTech
Pfizer/BioNTech admin-
administration
istration of the first dose on day 0 and the second dose on day 21. The antibody response is reported as a log 2 fold-change
of the first dose on day 0 and the second dose on day 21. The antibody response is reported as a log2 fold-change compared
compared to day 0. A positive response was arbitrarily scored for a log2 fold-change >1.5. (b) Summary of IgG
to day 0. A positive response was arbitrarily scored for a log2 fold-change >1.5. (b) Summary of IgG sero-reactivities shown
sero-reactivities shown in panel (a).
in panel (a).

Reactivity
Reactivity to Pep2_Spike,
to Pep2_Spike, corresponding
corresponding to Spiketo 287–317
Spike287–317
, was, observed
was observed in the
in the sera fromsera
from all vaccinated subjects, except vax_12, vax_14, and vax_17,
all vaccinated subjects, except vax_12, vax_14, and vax_17, in which antibodies against this in which antibodies
against
peptide thisunder
were peptide thewere underthreshold
positivity the positivity
during threshold
the entire during
study the entire
period. study period.
However, the
However,
antibody the antibody
response against response
Pep2_Spike against
showed Pep2_Spike showed different
different dynamics dynamics
in different subjects. inIndif-
ferent
fact, many subjects. In fact,
participants many participants
resulted negative to the resulted
detection negative to the detection
of antibodies of antibodies
against this peptide
at against
day 15 this
afterpeptide
the firstatdose.
day 15 Inafter the first
addition, dose. In addition,
anti-Pep2_Spike anti-Pep2_Spike
antibodies antibodies
in the vaccinated
in the vaccinated subjects vax_1 and vax_7 are specifically detected
subjects vax_1 and vax_7 are specifically detected as early as 15 days after the first dose, as early as 15but days
decreased under the threshold in sera sampled at day 60, i.e., 39 days after the second dose.39
after the first dose, but decreased under the threshold in sera sampled at day 60, i.e.,
days after
Positivity the second dose.
to Pep5_Spike, PositivitytotoSpike
corresponding Pep5_Spike,
802–819 , was corresponding
scored only for to aSpike 802–819, was
few subjects,
scored
while manyonly for a few subjects,
participants while many
never displayed participants
a detectable never response
antibody displayedtoa this
detectable
peptidean-
fortibody response
the entire periodto this peptide Pep6_Spike
of observation. for the entire (Spike 601–640 )ofhad
period observation. Pep6_Spike
the widest reactivity
(Spikeamong
pattern 601–640) had the widest reactivity pattern among the vaccinated subjects, with only
the vaccinated subjects, with only sera from participants vax_13 and vax_17
sera from participants
not reacting to it. A similar vax_13 and vax_17
diversity not reacting
of antibody response to init. the
A similar diversity
participating of anti-
subjects
wasbody
alsoresponse
observable in toward
the participating
Pep10_Spike subjects 524–598
(Spikewas also observable
). Overall, toward Pep10_Spike
all vaccinated subjects,
(Spike
except 524–598). Overall,
participant vax_17,allshowed
vaccinated subjects,
specific except
antibodies to participant vax_17, at
at least one peptide showed
both 30specific
and
60antibodies
days afterto theat injection
least one ofpeptide at both
the first dose30 ofand
the 60 days after the injection
Pfizer/BioNTech vaccine.ofConversely,
the first dose
of thevax_17
subject did not mount
Pfizer/BioNTech an antibody
vaccine. Conversely, response
subject against
vax_17 any didofnot
the mount
peptides anused in
antibody
theresponse
assay foragainst
the entireanyperiod
of the of observation.
peptides used in Indeed,
the assayvax_17for scored negative
the entire period inofanobserva-
assay
performed at day
tion. Indeed, 60 using
vax_17 the mixed
scored peptides
negative as the performed
in an assay immuno-adsorbent at day 60phase
using(Figure
the mixed2).
Specifically,
peptides as in the
thisimmuno-adsorbent
last assay, participant vax_8
phase also scored
(Figure below the in
2). Specifically, positivity
this lastthreshold,
assay, par-
due to a past
ticipant history
vax_8 alsoof scored
COVID-19. In fact,
below the the levels ofthreshold,
positivity antibodiesdue weretoalready
a pastvery high of
history
Vaccines 2021, 9, 987 6 of 8
Vaccines 2021, 9, 987 6 of 8
Vaccines 2021, 9, 987 5 of 7

COVID-19. In fact, the levels of antibodies were already very high in the serum of vax_8
COVID-19.
collected In fact,
at day the levelsthe
0; therefore, of antibodies
vaccinationwere already
did not very
result in ahigh
largeinvariation
the serumonofday
vax_8
60
collected
over day
in the at day 0;
0 in terms
serum therefore,
of vax_8 the
of fold-change. vaccination did not result in a large variation on day 60
collected at day 0; therefore, the vaccination did not result in a large
over day 0on
variation inday
terms
60 of fold-change.
over day 0 in terms of fold-change.
Reactivity to pep-mix Spike
32 Reactivity to pep-mix Spike
32

16

16

60days
) 2)
8
(log2(log

8 60days
Change

4
Change

4
FoldFold

0.5
1

10 10

11 11

12 12

13 13

14 14

15 15

16 16

17 17

18 18

19 19
x_

x_

x_

x_

x_

x_

x_

x_

x_
0.5

_
va

va

va

va

va

va

va

va

va

ax

va vax

va vax

va vax

va vax

va vax

va vax

va vax

va vax

va vax
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1

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x_

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va

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va Samples

va
Samples

Figure
Figure2.2.IgG
IgGimmunoreacting
immunoreacting60 60days
daysafter
afteradministration
administrationof
ofthe
thefirst
firstdose
doseofof the
the Pfizer/BioNTech
Pfizer/BioNTech
Figure 2.using
vaccine IgG immunoreacting
the mixed peptides 60 days after
as the administration ofphase.
immuno-adsorbent the firstThe
dose of the Pfizer/BioNTech
antibody response is re-
vaccine using the mixed peptides as the immuno-adsorbent phase. The antibody response is reported
vaccineasusing
ported a log2the mixed peptides
fold-change as the
of ELISA immuno-adsorbent
absorbance phase.
at day 60 with Thetoantibody
respect day 0, whenresponse is re-
all partic-
as a log2 fold-change of ELISA absorbance at day 60 with respect to day 0, when all participants,
ported except
ipants, as a log 2 fold-change
vax_8, of ELISA absorbance
were seronegative at day 60antibodies.
to anti-SARS-CoV-2 with respect to day 0, when all partic-
ipants, vax_8, were
except except vax_8,seronegative to anti-SARS-CoV-2
were seronegative antibodies.
to anti-SARS-CoV-2 antibodies.
To verify the reliability of the peptide-based ELISA test, we monitored the presence
To verify the reliability of the peptide-based ELISA test, we monitored the presence
To verify the reliability
ofofneutralizing of the peptide-based ELISA of test, we monitored the presence
neutralizingantibodies
antibodiestotoSARS-CoV-2
SARS-CoV-2in inthe
theserum
serum of vaccinated
vaccinatedsubjects
subjectsusing
usinganan
of neutralizing test.
FDA-approved antibodies
As to SARS-CoV-2
shown in Figure 3, in
the the serum
sera of allofvaccinated
vaccinatedsubjects
subjectsat using
30 daysan
FDA-approved test. As shown in Figure 3, the sera of all vaccinated subjects at 30 days (T1)
FDA-approved
(T1) test. As shown in Figure 3, the sera of all vaccinated ansubjects at neutral-
30 days
andand
at 60atdays
60 days
(T2)(T2)
afterafter injection
injection of the
of the firstfirst
dosedose
(T0)(T0) showed
showed effective
an effective neutralizing
(T1)
izing and at 60
capacity, days
with (T2) after
the exceptioninjection of the
of subject first dose (T0) showed an effective neutral-
capacity, with the exception of subject vax_17.vax_17. Interestingly,
Interestingly, in two patients,
in two patients, we observedwe
izing capacity,
observed a with
partial the exception
reduction in the of subject vax_17.
percentage of Interestingly,
inhibition 60 days in twothe
after patients,
first we
dose,
a partial reduction in the percentage of inhibition 60 days after the first dose, suggesting
observed
suggesting a partial reduction
that monitoring in the
SARS-CoV-2 percentage of inhibition
neutralization over 60 days after
time byresponse the first
antibodycould dose,
response
that monitoring SARS-CoV-2 neutralization over time by antibody help
suggesting
could help that
in monitoring
defining the SARS-CoV-2
level of neutralization
individual protection, over
as timeas
well bytheantibody
herd response
immunity
in defining the level of individual protection, as well as the herd immunity achieved by a
could
achieved help
byin
population. defining the level of individual protection, as well as the herd immunity
a population.
achieved by a population.
SARS-CoV-2 Neutralization Antibody Detection
110 SARS-CoV-2 Neutralization Antibody Detection
110
100
100
90
90
(%) (%)

80
of Inhibition

80
70
of Inhibition

70
60
60
50
Percentage

50
Percentage

40
40
30
30
20
T0
20
10 T0(30days)
T1
10 T2
T1(60days)
(30days)
0
T2 (60days)
0
C LC−T L −
C LC−T L −
va _1va 1
va 2va 2
va _3va 3
va 4va 4
va 5va 5
va _6va 6
va 7va 7
va _8va 8
va _v9a _9
va 1v0a 10
va _1v1a 11
va 1v2a 12
va _1v3a 13
va 1v4a 14
va _1v5a 15
va 1v6a 16
va 1v7a 17
va _1v8a 18
C _C19T 19

C LC+T +
L L+
x x_
x_ x_
x x_
x_ x_
x_ x_
x x_
x_ x_
x x_

L
x x
x_ x_
x x_
x_ x_
x x_
x_ x_
x x_
x_ x_
x_ x_
x x_
x x_
TR R
TR R
TR R
TR R
va

+
va

Figure 3. Qualitative direct detection of total neutralizing antibodies to SARS-CoV-2 at day 0 (T0),
Figure 3. Qualitative direct detection of total neutralizing antibodies to SARS-CoV-2 at day 0 (T0),
day30
30(T1),
Figure
day (T1),
3. andday
day60
Qualitative
and 60 (T2)detection
direct
(T2) afteradministration
after administration ofthe
thefirst
first
of total neutralizing
of doseof
ofPfizer/BioNTech.
antibodies
dose Pfizer/BioNTech.
to SARS-CoV-2 at day 0 (T0),
day 30 (T1), and day 60 (T2) after administration of the first dose of Pfizer/BioNTech.
Vaccines 2021, 9, 987 6 of 7

Using an ELISA based on synthetic peptides derived from the spike protein in the
immuno-adsorbent phase, we showed that the antibody response elicited by inoculation of
the two doses of the Pfizer/BioNTech vaccine appears to be qualitatively individual, both in
the type of peptides seen and in the temporal persistence of the antibody response. Overall,
all vaccinated subjects except one showed reactive antibodies to at least one peptide at both
30 and 60 days after the injection of the first dose of the Pfizer/BioNTech vaccine. Only
one of the 19 analyzed subjects showed no antibody response toward any of the selected
peptides, either suggesting a reduced humoral response to the Pfizer/BioNTech vaccine
or indicating that further epitopes from the spike proteins could be immunodominant.
However, the modest antibody response found in subject vax_17 was confirmed by the
poor neutralizing capacity found in its serum at different time points. One caveat to keep
in mind is that the antibody positivity threshold was arbitrarily set at a value of log2 >1.5
which corresponds to a ~2.8 linear increase in the absorbance value measured by the ELISA.
Overall, this short report shows results that are consistent with previously published data,
indicating that SARS-CoV-2 mRNA vaccination induces diverse antibodies toward the
spike protein [9,10], and that a single dose of mRNA vaccine is enough to elicit rapid
humoral responses in seropositive participants [11]. In addition, we observed that in a few
subjects, the antibody reactivity to spike’s peptides was reduced at 60 days after inoculation
of the first dose, that is, 39 days after injection of the second dose. This means that for
effective pandemic control, it is important to constantly monitor the antibody protection in
the population, and the assay described here could be a valid tool for this purpose.
An intriguing aspect to further investigate is the T cell-mediated response to syn-
thetic peptides used in this study. Since the beginning of the pandemic, seroconversion
in convalescent patients has been extensively monitored due to the technical ease of spe-
cific antibody detection. Nonetheless, although the analysis of T cell immunity may be
more challenging to carry out on a population-wide scale, it is crucial for patients who
do not mount an efficacious antibody response toward virus infection and/or vaccina-
tion [12,13]. More specifically, it would be interesting to evaluate whether the synthetic
peptides described here and in our previous work could recapitulate, or partially represent,
the portfolio of viral epitopes presented to T cells after both infection and vaccination,
and if any reactivity might be informative of a long-term effective immune protection.
In addition, they could also be useful for evaluating potential T cell cross-reactivity in
unexposed individuals due to endemic diffusion of cold coronaviruses [12].
From such a perspective, the development of synthetic peptide-based clinical tests
aimed to measure T cell immunity may also guide physicians in determining, on the one
hand, when booster vaccination is appropriate and, on the other hand, whether vaccination
strategies are able to contain the spread of variant forms of SARS-CoV-2 [13,14]. Overall,
collecting data on the ability of both antibodies and T lymphocytes to recognize SARS-CoV-
2 peptides from convalescent and/or vaccinated individuals would allow to obtain more
consistent and complete information on long-term protection after the ongoing vaccination
campaigns and toward newly recognized variants of concern [14].

Author Contributions: Conceptualization, P.V., T.Z. and I.P.; methodology, I.P. and L.Z.; validation,
I.P., A.P. and G.C.; resources, R.S. and P.V.; data curation, S.V., R.V. and J.R.M.; writing—original draft
preparation, P.V.; writing—review and editing, T.Z. and L.Z.; funding acquisition, P.V. All authors
have read and agreed to the published version of the manuscript.
Funding: This research was partially funded by a INBIOMED PON RI 2014-2020-MIUR-CUP
F26C18000160005 grant.
Institutional Review Board Statement: This study was approved by the Institutional Review Board
of Consorzio Sannio Tech (n. 01/2021).
Informed Consent Statement: Written informed consent was obtained from all subjects involved in
the study.
Data Availability Statement: Data are available on request from the corresponding authors.
Vaccines 2021, 9, 987 7 of 7

Acknowledgments: The authors thank Natalia Olivieri and Piero Porcaro for technical and logistic support.
Conflicts of Interest: The authors declare no conflict of interest.

Abbreviations

COVID-19 Coronavirus disease 2019

mRNA Messenger ribonucleic acid
SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2
ELISA Enzyme-linked immunosorbent assay

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