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1.Mulligan MJ, L yke KE, Kitchin N, et al. Phase 1/2 study  of COVI D-19 RNA vaccine 
BNT162b1 in adults. Nature. 2020;10.1038/s41586 -020-2639 -4.
2.Walsh EE, Frenck FW, Falsey  AR, et al. Safety and immunogenicity  of two RNA -based 
Covid -19 vaccine candidates. N Engl J Med 2020; DOI : 10.1056/NEJMoa2027906.
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Phase 1/2 study of COVID-19 RNA vaccine 
BNT162b1 in adults
Mark J. Mulligan, Kirsten E. Lyke, Nicholas Kitchin, Judith Absalon, Alejandra Gu tman, 
Stephen Lockhart, Kathleen Neuzil, Vanessa Raabe, Ruth Bailey, Kena A. Swanson, Ping Li, Kenneth Koury, Warren Kalina, David Cooper, Camila Fontes-Garfias  Pei-Yong Shi,  
Özlem Türeci, Kristin R. Tompkins, Edward E. Walsh, Robert Frenck  Ann R  Falsey,  
Philip R. Dormitzer, William C. Gruber, Uğur Şahin & Kathrin U  Jansen
This is a PDF file of a peer-reviewed paper that has been accepted for publication. 
Although unedited, the content has been subjected to preliminary formatting. Nature is providing this early version of the typese  paper as a service to our authors and readers. The text and figures will undergo copyediting and a proof review before the paper is published in its final form  Please note that during the production process errors may be discovered which ould affect the content, and all legal disclaimers apply.Received: 29 June 2020
Accepted: 4 August 2020Accelerated Article Preview Published 
online 12 August 2020
Cite this article as: Mulligan, M. J. et al. 
Phase 1/2 study of COVID-19 RNA vaccine BNT162b1 in adults. Nature https://doi.
org/10.1038/s41586-020-2639-4 (2020).https://doi.org/10.1038/s41586-020-2639-4
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Nature  | www.nature.com  | 1
ArticlePhase 1/2 study of COVID-19 RNA vaccine 
BNT162b1 in adults
Mark J. Mulligan1,2,12, Kirsten E. Lyke3,12, Nicholas Kitchin4,12, Judith Absalon5 ✉,  
Alejandra Gurtman5, Stephen Lockhart4, Kathleen Neuzil3, Vanessa Raabe1,2, Ruth Bailey4, 
Kena A. Swanson5, Ping Li6, Kenneth Koury5, Warren Kalina5, David Cooper5,  
Camila Fontes-Garfias7, Pei-Yong Shi7, Özlem Türeci8, Kristin R. Tompkins5,  
Edward E. Walsh9,10, Robert Frenck11, Ann R. Falsey9,10, Philip R. Dormitzer5, William C. Gruber5, 
Uğur Şahin8 & Kathrin U. Jansen5
In March 2020, the World Health Organization (WHO) declared a pandemic of 
coronavirus disease 2019 (COVID-19), due to severe acute respi atory syndrome coronavirus 2 (SARS-CoV-2)
1. With rapidly accumulating cases and deaths reported 
globally2, a vaccine is urgently needed. We report the a ailable safety, tolerability, and 
immunogenicity data from an ongoing placebo-controlled, observer-blinded dose escalation study among 45 healthy adults, 8 to 55 years of age, randomized to receive 2 doses, separated by 21 days, of 10 µg, 30 µg, or 100 µg of BNT162b1, a lipid nanoparticle-formulated, nucleoside-modified mRNA vaccine that encodes trimerized SARS-CoV-2 spike glycoprotein eceptor-binding domain (RBD). Local reactions and systemic events were dose-dependent, generally mild to moderate, and transient. A second vaccination with 100 µg was not administered due to increased reactogenicity and a lack of meaningfully increased immunogenicity after a single dose compared to the 30 
µg dose. RBD-binding IgG concentrations and SARS-CoV-2 
neutralizing titers in era increased with dose level and after a second dose. Geometric mean neutral zing titers reached 1.9- to 4.6-fold that of a panel of COVID-19 convalescent human sera at least 14 days after a positive SARS-CoV-2 PCR. Thes
e 
results support further evaluation of this mRNA vaccine candidate. (ClinicalTrials.gov identifier: NCT04368728).
In December 2019, a pneumonia outbreak f unknown cause occurred 
in Wuhan, China. By January 2020, a novel coronavirus was identified as 
the etiologic agent. Within a month  the genetic sequence of the virus 
became available (MN908947.3). Severe ac te respiratory syndrome 
coronavirus 2 (SARS-CoV-2) infec ions and the resulting disease, coro-
navirus disease 2019 (COVID-19), have spread globally. On 11 March 
2020, the World Health Organization (WHO) declared the COVID-19 
outbreak a pandemi1. To date, the United States has reported the 
most cases globally3. No vaccines are currently available to prevent 
SARS-CoV-2 infe tion or COVID-19.
The RNA vaccine platform has enabled rapid vaccine development 
in response to this pandemic. RNA vaccines provide flexibility in the 
design and expression of vaccine antigens that can mimic antigen 
structure and expression during natural infection. RNA is required 
for protein s nthesis, does not integrate into the genome, is transiently 
expressed, and is metabolized and eliminated by the body’s natural 
mechanisms and, therefore, is considered safe4–7. RNA-based prophylac -
tic infectious disease vaccines and RNA therapeutics have been shown 
o be safe and well-tolerated in clinical trials. In general, vaccination with RNA elicits a robust innate immune response. RNA directs expression of the vaccine antigen in host cells and has intrinsic adjuvant effects
8. 
A strength of the RNA vaccine manufacturing platform, irrespective of 
the encoded pathogen antigen, is the ability to rapidly produce large quantities of vaccine doses against a new pathogen
9,10.
Vaccine RNA can be modified by incorporating 1-methyl-pseudouridine  
which dampens innate immune sensing and increases mRNA translation 
in vivo11. The BNT162b1 vaccine candidate now being studied clinically 
incorporates such nucleoside-modified messenger RNA (modRNA) 
and encodes the receptor-binding domain (RBD) of the SARS-CoV-2 
spike protein, a key target of virus-neutralizing antibodies12–14. The 
RBD antigen expressed by BNT162b1 is modified by the addition of a 
T4 fibritin-derived foldon trimerization domain to increase its immu -
nogenicity15 by multivalent display16. The proper folding of the RBDs in 
the resulting protein construct has been confirmed by high resolution 
structural analysis (U.S., manuscript in preparation)17. The vaccine RNA 
is formulated in lipid nanoparticles (LNPs) for more efficient delivery 
into cells after intramuscular injection18. BNT162b1 is one of several 
RNA-based SARS-CoV-2 vaccine candidates being studied in parallel https://doi.org/10.1038/s41586-020-2639-4
Received: 29 June 2020Accepted: 4 August 2020Published online: 12 August 2020
1New York University Langone Vaccine Center, New York, NY, USA. 2New York University Grossman School of Medicine, New York, NY, USA. 3University of Maryland School of Medicine, Center 
for Vaccine Development and Global Health, Baltimore, MD, USA. 4Vaccine Research and Development, Pfizer Inc, Hurley, UK. 5Vaccine Research and Development, Pfizer Inc, Pearl River, NY, 
USA. 6Vaccine Research and Development, Pfizer Inc, Collegeville, PA, USA. 7University of Texas Medical Branch, Galveston, TX, USA. 8BioNTech, Mainz, Germany. 9University of Rochester, 
Rochester, NY, USA. 10Rochester General Hospital, Rochester, NY, USA. 11Cincinnati Children’s Hospital, Cincinnati, OH, USA. 12These authors contributed equally: Mark J. Mulligan, Kirsten E. 
Lyke, Nicholas Kitchin. ✉e-mail: [email protected] ARTICLE PREVIEW 
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Articlefor selection to advance to a safety and efficacy trial. Here, we present 
available data, through 14 days after a second dose in adults 18 to 55 
years of age, from an ongoing Phase 1/2 vaccine study with BNT162b1, 
which is also enrolling adults 65 to 85 years of age (ClinicalTrials.gov 
identifier: NCT04368728).
Study Design and Demographics
Between 04 May 2020 and 19 June 2020, 76 participants were screened, 
and 45 participants were randomized and vaccinated. Twelve partici-
pants per dose level (10 µg and 30 µg) were vaccinated with BNT162b1 
on Days 1 and 21, and 12 participants received a 100- µg dose on Day 1. 
Nine participants received placebo (Figure 1 ). The study population 
consisted of healthy male and nonpregnant female participants with a 
mean age of 35.4 years (range: 19 to 54 years); 51.1% were male and 48.9% 
were female. Most participants were white (82.2%) and non-Hispanic/
non-Latinx (93.3%) (Extended Data Table 1).
Safety and T olerability
In the 7 days following either Dose 1 or 2, pain at the injection site was 
the most frequent solicited local reaction, reported after Dose 1 by 
58.3% (7/12) in the 10-µg, 100.0% (12/12 each) in the 30- µg and 100-µg 
BNT162b1 groups, and 22.2% (2/9) in the placebo group. After Dose 2, 
pain was reported by 83.3% (10/12) and 100.0% of BNT162b1 recipients 
at the 10- µg and 30- µg dose levels, respectively, and by 16.7% of pla -
cebo recipients. All local reactions were mild or moderate in severity 
except for one report of severe pain following Dose 1 of 100 µg BNT162b1  
(Figure  2; Extended Data Table 2).
The most common systemic events reported in the 7 days after each 
vaccination in both BNT162b1 and placebo recipients were mild to 
moderate fatigue and headache. Reports of fatigue and headache were 
more common in the BNT162b1 groups compared to the placebo group. 
Additionally, chills, muscle pain, and joint pain were reported among 
BNT162b1 recipients and not in placebo recipients. Systemic eve ts 
increased with dose level and were reported in a greater number of 
participants after the second dose (10-µg and 30-µg groups). Follow -
ing Dose 1, fever (defined as ≥38.0 °C) was reported by 8.3% (1/12) of 
participants in both the 10-µg and 30-µg groups and by 50.0% (6/12) of 
BNT162b1 recipients in the 100-µg group. Following Dose 2  8.3% (1/12) 
of participants in the 10-µg group and 75.0% (9/12) of participants in 
the 30-µg group reported fever ≥38.0 °C  Based on the reactogenicity 
reported after the first dose of 100 µg and the second dose of 30 µg, 
participants who received an initial 100- µg dose did not receive a sec-
ond 100- µg dose. Fevers generally resolved within 1 day of onset. No 
Grade 4 systemic events or fever were r ported. (Figure  3a, b , Extended 
Data Table 3). Most local reactions and systemic events peaked by Day 
2 after vaccination and resolved by Day 7.
Adverse events (AEs  (Extended Data Table 4) were reported by 50.0% 
(6/12) of participants who received either 10 µg or 30 µg of BNT162b1, 58.3% (7/12) of those who received 100 µg of BNT162b1, and 11.1% (1/9) of placebo recipients. Two participants reported a severe AE: Grade 3 
fever 2 day  after vaccination in the 30-µg group, and sleep disturbance 
1 day after vaccination in the 100-µg group. Related AEs were reported 
by 2 % (3/12 n the 10-µg groups) to 50% (6/12 each in 30-µg and 100-µg 
groups) of BNT162b1 recipients and by 11.1% (1/9) of placebo recipients. 
No serious adverse events (SAEs) were reported.
No Grade 1 or greater change in routine clinical laboratory values or 
laboratory abnormalities were observed for most participants after 
either of the BNT162b1 vaccinations. Of those with laboratory changes, 
the largest changes were decreases in lymphocyte count after Dose 1 
in 8.3% (1/12), 45.5% (5/11), and 50.0% (6/12) of 10 µg, 30 µg, and 100 µg 
BNT162b1 recipients, respectively. One participant each in the 10-µg 
group (8.3% [1/12]) and 30-µg group (9.1% [1/11]) dose levels and 4 par-
ticipants in the 100-µg group (33.3% [4/12]) had Grade 3 decreases in lymphocytes. These post-Dose 1 decreases in lymphocyte count, 
were transient and returned to normal 6 to 8 days after vaccination 
(Extended Data Figure 1). In addition, Grade 2 neutropenia was noted 6 to 8 days after the second dose in 1 participant each in the 10-µg and 
30-µg BNT162b1 groups. These two participants continue to be followed 
in the study and no AEs or clinical manifestations of neutropenia have 
been reported to date. None of the postvaccination abnormalities 
observed were associated with clinical findings.
Immunogenicity
RBD-binding IgG concentrations and SARS-CoV-2 neutralizing titers 
were assessed at baseline, and at 7 and 21 days after the first dose  and 
at 7 (Day 28) and 14 days (Day 35) after the second dose of BNT162b1. By  
21 days after the first dose (for all three dose levels)  geometric mean concentrations (GMCs) of RBD-binding IgG ranged from 534 to 1,778 U/mL (Figure  4a). In comparison, a panel of 38 SARS-CoV-2 infection/
COVID-19 convalescent sera drawn at least 14 days after a polymer -
ase chain reaction (PCR)-confirmed diagnosis from patients 18 to 83 
years of age had an RBD-binding IgG GMC of 602 U/mL. (Additional 
information on the convalescent serum panel is presented in Meth -
ods.) By 7 days after the second dose (for the 10 µg and 30 µg dose 
levels) RBD-binding IgG GMCs had increased to 4,813 to 27,872 U/mL.  
RBD-binding antibody concentrations among participants who 
received one dose of 100 µg BNT162b1 did not increase beyond 21 days 
after the first vaccination. In the participants who received the 10 µg 
and 30 µg doses of BNT162b1, highly elevated RBD-binding antibody 
concentrat ons persisted to the last time point evaluated (Day 35, 14 
days af er the second dose). These RBD-binding antibody concentra-tions were 5,880 to 16,166 U/mL compared to 602 U/mL in the human conv lescent serum panel.
For al  doses, small increases in SARS-CoV-2 neutralizing geometric 
mean titers (GMTs) were observed 21 days after Dose 1 (Figure 4b ). 
Substantially greater serum neutralizing GMTs were achieved 7 days 
fter the second 10 µg and 30 µg dose, reaching 168 to 267. Neutralizing 
GMTs further increased by 14 days after the second dose to 180 at the 
10 µg dose level and 437 at the 30 µg dose level, compared to 94 for the 
convalescent serum panel. The kinetics and durability of neutralizing titers are being monitored.
Discussion
The RNA-based SARS-CoV-2 vaccine candidate BNT162b1 administered 
at 10 µg, 30 µg, and 100 µg to healthy adults 18 to 55 years of age exhib -
ited a tolerability and safety profile consistent with those previously 
observed for mRNA-based vaccines5. A clear dose-level response in 
elicited neutralizing titers was observed after Doses 1 and 2 in adults 
18 to 55 years of age with a particularly steep dose response between 
the 10 µg and 30 µg dose levels.
Based on the tolerability profile of the first dose at 100 µg and the 
second dose at 30 µg, participants randomized to the 100-µg group 
did not receive a second vaccination. Reactogenicity was generally 
greater after the second dose in the other two dosing levels; however, 
symptoms were transient and resolved within a few days. Transient 
decreases in lymphocytes (Grades 1-3) were observed within a few 
days after vaccination, with lymphocyte counts returning to baseline 
within 6 to 8 days in all participants. These laboratory abnormalities 
were not associated with clinical findings. RNA vaccines are known 
to induce type I interferon, which has been associated with transient 
migration of lymphocytes into tissues19–22.
Robust immunogenicity was observed after vaccination with 
BNT162b1. RBD-binding IgG concentrations were detected at 21 days 
after the first dose and substantially increased 7 days after the second 
dose given at Day 21. After the first dose, the RBD-binding IgG GMCs 
(10 µg dose recipients) were similar to those observed in a panel of  ACCELERATED ARTICLE PREVIEW 
   
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Nature  | www.nature.com  | 338 convalescent human serum samples, obtained at least 14 days after a 
PCR-confirmed diagnosis of SARS-CoV-2 infection/COVID-19. Post-Dose 
1 GMCs were similar in the 30 µg and 100 µg groups and higher than 
those in the convalescent serum panel. After Dose 2 with 10 µg or  
30 µg BNT162b1, the RBD-binding IgG GMCs were ~8.0-fold to ~50-fold 
that of the convalescent serum panel GMC.
The higher RBD-binding IgG GMC elicited by the vaccine relative 
to the GMC of the human convalescent serum panel may be attrib -
uted, in part, to antibodies that bind epitopes that are exposed on the 
RNA-expressed RBD immunogen and the recombinant RBD target 
antigen of the binding assay but are buried and inaccessible to antibody 
on the RBDs that are incorporated into the spikes of SARS-CoV-2 viri-
ons. Therefore, neutralization provides a measure of vaccine-elicited antibody response that is more relevant to potential protection. Neu-
tralization titers were measurable after a single vaccination at Day 21 for 
all dose levels. At Day 28 (7 days after Dose 2), substantial SARS-CoV-2 
neutralization titers were observed. The virus-neutralizing GMTs after 
the 10 µg and 30 µg Dose 2 were, respectively, 1.8-fold and 2.8-fold the 
GMT of the convalescent serum panel. By Day 35 (14 days after Dose 
2), despite the decrease in RBD-binding IgG titers since Day 28, neu -
tralizing GMTs continued to rise, to 1.9-fold and 4.6-fold the GMT of 
the convalescent panel for the 10 µg and 30 µg doses, respectively, 
consistent with affinity maturation.
Assuming that neutralization titers induced by natural infection 
provide protection from COVID-19 disease, comparing vaccine-induced 
SARS-CoV-2 neutralization titers to those from sera of convalescent 
humans provides a benchmark for the magnitude of the vaccine-elicited 
response and the vaccine’s potential to provide protection. Because 
a protective human neutralizing titer is unknown, these findings are 
not proof of vaccine efficacy. Efficacy will be determined in a pivotal 
Phase 3 trial. Because the 100 µg dose level cohort was not boosted, 
no data for immunogenicity after a second vaccination at this dose 
level are available; however, there were no substantial differences in 
immunogenicity between the 30 µg and 100 µg dose levels after Dose 
1. This observation suggests that a well-tolerated and immunoge ic 
dose level may be between 10 µg and 30 µg for this vaccine candidate.
Our study had several limitations. While we used convalescent sera 
as a comparator, the kind of immunity (T cells versu  B cells or both) 
and level of immunity needed to protect from COVID-19 are unknown. 
Further, this analysis of available data did not assess immune responses 
or safety beyond 2 weeks after the second dose of vaccine. Both are 
important to inform the public health use of this vaccine. Follow-up 
will continue for all participants and will include collection of SAEs for  
6 months and COVID-19 infection and multiple additional immuno -
genicity measurements through up to two years. While our population 
of healthy adults 55 years of age and younger is appropriate for a Phase 
1/2 study, it does not accurat ly re lect the population at highest risk for 
COVID-19. Adults 65 years of ag  and over have already been enrolled in 
this study and results will be reported as they become available. Later 
phases of this study will prioritize enrollment of more diverse popula -
tions, including hose with chronic underlying health conditions and from racial/ethnic groups adversely affected by COVID-19
23.
The clin cal testing of BNT162b1 described here has taken place in the 
context of a b oader, ongoing COVID-19 vaccine development program. 
That program includes the clinical testing of three additional vaccine 
cand dates  ncluding candidates encoding the full-length spike, and a 
parallel trial in Germany, in which additional immune responses, includ -
ing neutralizing responses against variant strains and cell-mediated 
responses, are being assessed (U.S. manuscript in preparation)24. The 
resulting comparative data will allow us to address whether a full-length 
spike immunogen, which presents additional epitopes, is better able 
than the relatively small RBD immunogen encoded by BNT162b1 to 
elicit high virus neutralizing titers that are robust to potential antigenic drift of SARS-CoV-2. The clinical findings for the BNT162b1 RNA-based 
vaccine candidate are encouraging and strongly support accelerated 
clinical development, including efficacy testing, and at-risk manu -
facturing to maximize the opportunity for the rapid production of a 
SARS-CoV-2 vaccine to prevent COVID-19.
Online content
Any methods, additional references, Nature Research reporting sum-
maries, source data, extended data, supplementary information, 
acknowledgements, peer review information; details of author con -
tributions and competing interests; and statements of data and code 
availability are available at https://doi.org/10.1038/s41586 020 2639-4 .
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Figure 1 | Disposition of participants.  Participants not assigned (n=20) were screened but not randomized because enrollmen  had closed.
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Figure 2 | Local reactions reported within 7 days of vaccination for all dose 
levels.  Solicited injection-site (local) reactions were: pain at injection site 
(mild: does not interfere with activity; moderate: interferes with activity; severe: prevents daily activity; Grade 4: emergency room visit or hospitalization) and redness and swelling (mild: 2.0 to 5.0 cm in diameter; moderate: >5.0 to 10.0 cm in diameter; severe: >10.0 cm in diameter; Grade 4: necrosis or exfoliative de mat tis for redness, and necrosis for swelling). Data were collected with the use of electronic diaries for 7 days after each vaccination.
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Figure 3 | a. Systemic even s and medication use reported within 7 days 
after Vaccination 1 for all dos  levels and b. After Vaccination 2 for the 10-µg and 30-µg dos  level . Solicited systemic events were: fatigue, headache, chills, new or orsened muscle pain, new or worsened joint pain (mild: does not nterfe e with activity; moderate: some interference with activ y; severe: pr vents daily activity), vomiting (mild: 1 to 2 times in 24 hours; mod rate: >2 t mes in 24 hours; severe: requires intravenous hydration), diarrhea (mild: 2 to 3 loose stools in 24 hours; moderate: 4 to 5 loose stools in  24 hours; severe: 6 or more loose stools in 24 hours); Grade 4 for all events: emergency room visit or hospitalization; and fever (mild: 38.0 °C to 38.4 °C; moderate: 38.5 °C to 38.9 °C; severe: 39.0 °C to 40.0 °C; Grade 4: >40.0 °C). Medication: proportion of participants reporting use of antipyretic or pain medication. Data were collected with the use of electronic diaries for 7 days after each vaccination.
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Figure 4 | Immunogenicity of BNT162b1. Participants in groups of 15 were 
vaccinated with the indicated dose levels of BNT162b1 (n=12) or with placebo (n=3) on Days 1 (all dose levels and placebo) and 21 (10 µg and 30 µg dose levels and placebo). Reponses in placebo recipients for each of the osing groups are combined. The 28-day bleed is 7 days after the second vaccin tion. Se a were obtained before vaccination (Day 1) and 7, 21, and 28 days after the fi st vaccination. Human COVID-19 convalescent sera (HCS, n=38) were obtained at least 14 days after PCR-confirmed diagnosis and t a tim  when the donors were asymptomatic. a. GMCs of recombinant RBD-binding IgG. Because Luminex 
assay measured antibody concentrations are in arbitrary units, they cannot be directly translated into concentrations on a molar or mass basis. Lower limit of quantitation is 1.15. b . 50% SARS-CoV-2 neutralizing GMTs. Each data point 
represents a serum sample, and each vertical bar represents a geometric mean with 95% CI. The number above the bars are either the GMC or GMT for the group. Arrows indicate timing of vaccination (blood draws were conducted prior to vaccination on vaccination days).
Nature  | www.nature.com  | 7   
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Methods
Study design
This study was conducted in healthy men and nonpregnant women  
18 to 55 years of age to assess the safety, tolerability, and immuno -
genicity of ascending dose levels of various BNT162 mRNA vaccine 
candidates. In the part of the study reported here, assessment of three 
dose levels (10- µg, 30- µg, or 100- µg) of the BNT162b1 candidate was 
conducted at two sites in the United States. This study utilized a sentinel 
cohort design with progression and dose escalation taking place after 
review of data from the sentinel cohort at each dose level.
Eligibility
Key exclusion criteria included individuals with known infection with 
human immunodeficiency virus, hepatitis C virus, or hepatitis B virus; 
immunocompromised individuals and those with a history of auto -
immune disease; and those with increased risk for severe COVID-19, 
previous clinical or microbiological diagnosis of COVID-19, receipt of 
medications intended to prevent COVID-19, previous vaccination with 
any coronavirus vaccine, a positive serological test for SARS-CoV-2 
IgM and/or IgG at the screening visit, and a SARS-CoV-2 nucleic acid 
amplification test (NAAT)-positive nasal swab within 24 hours before 
study vaccination.
The final protocol and informed consent document were approved 
by institutional review boards for each of the participating investiga-tional centers. This study was conducted in compliance with all Inter-
national Council for Harmonisation (ICH) Good Clinical Practice (GCP) 
guidelines and the ethical principles of the Declaration of Helsinki. 
A signed and dated informed consent form was required before any 
study-specific activity was performed.
Endpoints
In this report, results from the following study primary endpoints are 
presented: the proportion of participants reporting solicited local reac -
tions, systemic events, and use of antipyretic and/or pain medi ation 
within 7 days after vaccination, AEs and SAEs (available through up to ~45 days after Dose 1), and the proportion of participants with clin cal 
laboratory abnormalities 1 and 7 days after vaccinat on and grading 
shifts in laboratory assessments between baseline and 1 and 7 days after 
Dose 1 and between Dose 2 and 7 days after Dose 2  Secondary endpoints 
included: SARS-CoV-2 neutralizing GMTs and SARS CoV-2 RBD-binding 
IgG GMCs 7 and 21 days after Dose 1 and 7 and 14 days after Dose 2.
Procedures
Study participants were randomly assigned to a vaccine group using 
an interactive web-based response technology system with each group 
comprising 15 participants (12 ac ve vaccine recipients and 3 placebo 
recipients). Participants were to receive two 0.5-mL doses of either 
BNT162b1 or placebo  administered by intramuscular injection into 
the deltoid muscle.
BNT162b1 incorporates a Good Manufacturing Practice (GMP)-grade 
mRNA drug ubsta ce that encodes the trimerized SARS-CoV-2 spike 
glycoprotein RBD antigen. The coding sequence for the antigen has 
been deposited with GenBank, accession code MN908947.3. The mRNA 
is fo mulated with lipids as the mRNA-LNP drug product. The vaccine 
was supplied as a buffered-liquid solution for intramuscular injection 
and was stored at -80 °C. The placebo was a sterile saline solution for inject on (0.9% sodium chloride injection, in a 0.5-mL dose).
Safety assessments
Safety assessments included a 4-hour observation after vaccination 
(for the first 5 participants vaccinated in each group), or a 30-minute 
observation (for the remainder of participants) for immediate AEs. 
The safety assessments also included self-reporting of solicited local 
reactions (redness, swelling, and pain at the injection site), systemic events (fever, fatigue, headache, chills, vomiting, diarrhea, muscle 
pain, and joint pain), the use of antipyretic and/or pain medication in 
an electronic diary for 7 days after vaccination, and the reporting of 
unsolicited AEs and SAEs after vaccination. Hematology and chemistry 
assessments were conducted at screening, 1 and 7 days after Dose 1, 
and 7 days after Dose 2.
There were protocol-specified safety stopping rules for all sentinel 
cohort participants. Both an internal review committee and an external 
data monitoring committee reviewed all safety data. No stopping rules 
were met prior to the publication of this report.
Human convalescent serum panel
The 38 human SARS-CoV-2 infection/COVID-19 convalescent sera were 
drawn from participants 18 to 83 years of age, at least 14 days after 
PCR-confirmed diagnosis, and at a time when participants were asymp -
tomatic. The mean age of the donors was 45 years of age. Neutralizing 
GMTs in subgroups of the donors were as follows: ≤ 55 years of age 
- 82 (n=29); > 55 years of age – 142 (n=9); symptomatic infections – 90 
(n=35); asymptomatic infections – 156 (n=3)  The antibody titer for 
the one hospitalized individual was 618  The sera were obtained from 
Sanguine Biosciences (Sherman Oaks, CA), the MT Group (Van Nuys, 
CA), and Pfizer Occupational Health and Wellness (Pearl River, NY).
Immunogenicity assessments
50 mL of blood was colle ted for immunogenicity assessments before 
each study vaccination, at 7 and 21 days after Dose 1, and at 7 and 14 days 
after Dose 2  In the RBD-binding IgG assay, a recombinant SARS-CoV-2 
RBD contai ing a C terminal Avitag™ (Acro Biosystems Cat# SPD-C82E9) 
and no foldon domain was bound to streptavidin-coated Luminex® 
mic ospheres. Briefly, 1.25 × 107 microspheres/mL were coated with 
streptavidin by 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide 
hydrochloride (EDC) reaction. Recombinant RBD Avitag was coupled to 
s reptavidin beads by incubating for 90 minutes at room temperature 
with shaking (35 RPM). Beads were blocked in 1% BSA buffer for 30 min -
utes at room temperature. Heat-inactivated subject serum was diluted 
1:500, 1:5000, and 1:50000 in assay buffer (PBS with 0.5% BSA, 0.05% 
Tween, and 0.02% sodium azide). Following a 16- to 20-hour incubation 
at 2-8 °C with shaking (300 RPM), plates were washed three times in a 
solution containing 0.05% Tween-20. An R-Phycoerythrin-conjugated 
goat anti-human polyclonal antibody ( Jackson Labs) was then added 
to plates for 90 minutes at room temperature with shaking (300 RPM). 
Plates were then washed a final time in a solution containing 0.05% 
Tween-20. Data were captured as median fluorescent intensities using a 
Luminex reader and converted to U/mL antibody concentrations using 
a reference standard curve with arbitrary assigned concentrations 
of 100 U/mL and accounting for the serum dilution factor. The refer-
ence standard was composed of a pool of five COVID-19 convalescent 
serum samples (>14 days post PCR diagnosis). Three dilutions are used 
to increase the likelihood that at least one result for any sample will 
fall within the usable range of the standard curve. Assay results were 
reported in U/mL of IgG. The final assay results are expressed as the 
GMC of all sample dilutions that produced a valid assay result within 
the assay range.
The SARS-CoV-2 neutralization assay used a previously described 
strain of SARS-CoV-2 (USA_WA1/2020) that had been rescued by reverse 
genetics and engineered by the insertion of an mNeonGreen gene into 
open reading frame 7 of the viral genome25. This reporter virus gener-
ates similar plaque morphologies and indistinguishable growth curves 
from the wild-type virus. Viral master stocks (2 × 107 PFU/mL) used 
for the neutralization assay were grown in Vero E6 cells as previously 
described25. When testing patient convalescent serum specimens, the 
fluorescent neutralization assay produced comparable results as the 
conventional plaque reduction neutralization assay26. Briefly, serial 
dilutions of heat inactivated sera were incubated with the reporter 
virus to yield approximately a 10% to 30% infection rate of the Vero 
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monolayer) for 1 hour at 37 °C before inoculating Vero CCL81 cell mon -
olayers (targeted to have 8,000 to 15,000 cells per well) in 96 well plates 
to allow accurate quantification of infected cells. Total cell counts per 
well were enumerated by nuclear stain (Hoechst 33342) and fluorescent 
virally infected foci were detected 16 to 24 hours after inoculation with 
a Cytation™ 7 Cell Imaging Multi-Mode Reader (BioTek) with Gen5 Image 
Prime version 3.09. Titers were calculated in GraphPad Prism version 
8.4.2 by generating a 4-parameter (4PL) logistical fit of the percent 
neutralization at each serial serum dilution. The 50% neutralization 
titer (VNT50) was reported as the interpolated reciprocal of the dilution 
yielding a 50% reduction in fluorescent viral foci.
Statistical analysis
The sample size for the reported part of the study was not based on sta -
tistical hypothesis testing. The primary safety objective was evaluated 
by descriptive summary statistics for local reactions, systemic events, 
abnormal hematology and chemistry laboratory parameters, AEs, and 
SAEs after each vaccine dose for each vaccine group. The secondary 
immunogenicity objectives were descriptively summarized at the vari -
ous time points. All participants with data available were included in 
the safety and immunogenicity analyses.
Reporting summary
Further information on research design is available in the Nature 
Research Reporting Summary linked to this paper.
Data availability
Upon request, and subject to review, Pfizer will provide the data that 
support the findings of this study. Subject to certain criteria, condi -
tions, and exceptions, Pfizer may also provide access to the related 
individual anonymized participant data. See  https://www.pfizer.com/
science/clinical-trials/trial-data-and-results for more information. 
These data are interim data from an ongoing study, with the database 
not locked. Data have not yet been source verified or subjected to sta d -
ard quality check procedures that would occur at the time of database 
lock and may therefore be subject to change.
 
25. Xie X. et al. An infectious cDNA clone of SARS CoV-2. Cell Host Microbe. 27, 841-848 
(2020).
26
. Muruato, A.E., Fontes Garfias, C.R., Ren, P
. et al. A high throughput neutra izing antibody 
assay for COVID-19 diagnosis and vaccine evaluation. bioRxiv  https://doi.org/10.1101/  
2020.05.21.109546 (2020).Acknowledgements The authors would like to thank Carol Monahan and Deb Gantt (Pfizer 
Inc) for writing and editorial support and Hua Ma, James Trammel, and Kiran Challagali 
(Pfizer Inc) for statistical analysis support in the generation of this manuscript. We would like to thank all the participants who volunteered for this study. We also acknowledge the following individuals for their contributions to this work: NYU Langone Vaccine Center: Angelica Kottkamp, MD, Ramin Herati, MD, Rebecca Pellet Madan, MD, Mary Olson, DNP, ANP-BC, Marie Samanovic-Golden, PhD, Elisabeth Cohen, MD, Amber Cornelius, MS, Laura Frye, MPH, Heekoung Youn, RN, CCRC, MA, Baby Jane Fran, RN, Kanika Ballani, PharmD, MBA, Natalie Veling, RN, Juanita Erb, RN, BSN, MPA, Mahnoor Ali, BA, Lisa Zhao, BA, Stephanie Rettig, MPH, Hibah Khan, MPA, Harry Lambert, BA, Kelly Hu, BA, and Jonathan Hyde, BS. Staffing services were supported in part by an NYU CTSA grant (UL1 TR001445) from the National Center for Advancing Translational Sciences, National Institutes of Health. Center for Vaccine Development and Global Health, University of Maryland Sc ool of Medicine: Monica McArthur, MD, PhD, Justin Ortiz, MD, MS, FACP, FCCP, Rekha Rapaka, MD, Linda Wadsworth, RN, Ginny Cummings, RN, Toni Robinson, RN, Nancy G eenberg, RN, Lisa Chrisley, RN, Wanda Somrajit, RN, Jennifer Marron, RN, BSN, MS, Constanc  Thomas, RN, Kelly Brooks, RN, Lisa Turek, RN, Patricia Farley, RN, Staci Eddington  Pa agiot  Komninou, Mardi Reymann, Kathy Strauss, Biraj Shrestha, Sudhaunshu J shi, Robin Barnes, RN, Roohali Sukhavasi, Myounghee Lee, PharmD, Alyson Kwon, and Terry Sharp. University of Rochester and Rochester General Hospital: Emily Pierce, RN  and Mary Criddle, RN. Cincinnati Children’s Hospital: Amy Cline, RN, Susan Parker  RN, Michelle Di key, APRN, Kristen Buschle, APRN. Pfizer Inc: Andrea Cawein, John L  erez  MD, MSc, Harpreet Seehra, Dina Tresnan, DVM, PhD, Robert Maroko, MD, Helen Smith, Sarah Tweedy, Amy Jones, Greg Adams, Rabia Malick, Emily Worobetz, Erica Weaver  Lipi g Zhang  armel Devlin, Donna Boyce, Elisa Harkins Tull, Mark Boaz, Michael Cru , Vaccines Clinical Assay Team and Vaccines Assay Development Team. BioNTech: Corinna Rosenbaum, Christian Miculka, Andreas Kuhn, Ferdia Bates, Paul Strecker  nd A exandra Kemmer-Brück. BioNTech is the sponsor of the study. Pfizer was respons ble for he design, data collection, data analysis, data interpretation, and writing of the repo t  he corresponding authors had full access to all the data in the study and had final responsibility for the decision to submit the data for publication. All study data were available to all authors.
Author contributions KUJ, PRD, CG, NK  SL, AG, RB, and US were involved in the design of 
the overall study and strategy. KN, MJM, EEW, RF, and ARF provided feedback on the study design. WK, DC, KAS, KRT  CFG and PYS performed the immunological analyses. MJM, KN, EEW, RF, ARF, KEL, and VR olle ted data as study investigators. PL and KK developed the statistical desi n and oversaw the data analysis. JA, KUJ, PRD, and WCG drafted the initial version of the manuscript  All authors reviewed and edited the manuscript and approved the final version.
Compe ing interests NK, JA, AG, SL, RB, KAS, PL, KK, WK, DC, KRT, PRD, WCG, and KUJ are 
mployee  of Pfizer and may hold stock options. US and ÖT are stock owners, 
management board members, and employees at BioNTech SE (Mainz, Germany) and are 
nv tors on patents and patent applications related to RNA technology. MJM, KEL, KN, 
EEW, ARF, RF, and VR received compensation from Pfizer for their role as study 
nvestigators. CFG and PYS received compensation from Pfizer to perform the 
neutralization assay.
Additional information
Supplementary information is available for this paper at https://doi.org/10.1038/s41586-020-2639-4 .
Correspondence and requests for materials should be addressed to J.A.Peer review information Nature thanks Barbra Richardson and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.Reprints and permissions information is available at http://www.nature.com/reprints.
   
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Extended Data Figure 1 | Post Vaccination Changes in Lymphocyte Coun  
Over Time.  Figure represents box-and-whisker plots for obs rved values at the 
following timepoints: Dose 1/Day 1-3: ~1 day after Dose 1; Dose 2/Day 6 8: ~7 days after Dose 1; Pre-Dose 2: before Dose 2; Dose 2/Day 6-8:  7 days after Dose 2. Symbols denote group means – O: placebo; +: 10 µg; X: 30 µg; Δ: 100 µg. Center 
line of box denotes median; lower and upper edges denote first and third quartiles; lower and upper whiskers denote minimum and maximum.
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Extended Data Table 1 | Demographic Characteristics.
N = number of subjects in the specified group, or the total sample. This value is the denominator for e percen age calculations. n = Number of subjects with the specified characteristic.
   
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Extended Data Table 2 | Adverse Events.
N: number of subjects in the specified group or the total sample. This value is the denominator for the percentage calculations. n: number of subjects reporting at least 1 occurrence of the 
specified adverse event category. For "any event", n: the number of subjects reporting at least 1 occurr nce of any adverse event; Related: Assessed by the investigator as related to investiga-tional product.
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1 nature research  |  reporting summary April 2020
Corresponding author(s): Judith Absalon
Last updated by author(s): Jul 27, 2020
Reporting Summary
Nature Research wishes to improve the reproducibility of the work that we publish. This form provides structure for consistency and transparency 
in reporting. For further information on Nature Research policies, see our Editorial Policies  and the Editorial Policy Checklist .
Statistics
For all statistical analyses, confirm that the following items are present in the figure legend, table legend, main text, or Methods section.
n/a Confirmed
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A statement on whether measurements were taken from distinct samples or whether the same sample was measured repeatedly
The statistical test(s) used AND whether they are one  or two sided 
Only common tests should be described solely by name; describe more complex techniques in the Methods section.
A description of all covariates tested
A description of any assumptions or corrections, such as tests of normality and adjustment for multiple comparisons
A full description of the statistical parameters including central tendency (e.g. means) or other basic estimates (e.g. regression coefficient) AND variation (e.g. standard deviation) or associated estimates of uncertainty (e.g. confidence intervals)
For null hypothesis testing, the test statistic (e.g. F, t, r) with confidence intervals, effect sizes, degrees of freedom and P value noted 
Give P values as exact values whenever suitable.
For Bayesian analysis, information on the choice of priors and Markov chain Monte Carlo settings
For hierarchical and complex designs, identification of the appropriate level for tests and full reporting of outcomes
Estimates of effect sizes (e.g. Cohen's d, Pearson's r), indicating how they were calculated
Our web collection on statistics for biologists  contains articles on many of the points above.
Software and code
Policy information about availability of computer code
Data collection Inform (for data collected in the case report form) and electronic diary (Signant Health platform) for participant self reported reactogenicity
Data analysis SAS 9.4
For manuscripts utilizing custom algorithms or software that are central to the research but not yet described in published literature, software must be made available to editors and reviewers. We strongly encourage code deposition in a community repository (e.g. GitHub). See the Nature Research  guidelines for submitting code & software  for further information.
Data
Policy information about availability of data
All manuscripts must include a data availability statement . This statement should provide the following information, where applicable: 
 Accession codes, unique identifiers, or web links for publicly available datasets  A list of figures that have associated raw data  A description of any restrictions on data availability
 Upon request, and subject to review, Pfizer will provide the data that support the findings of this study. Subject to certain criteria, conditions and exceptions, Pfizer may also provide access to the related individual anonymized participant data. See https://www.pfizer.com/science/clinical trials/trial data and results for more information
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2 nature research  |  reporting summary April 2020Field-specific reporting
Please select the one below that is the best fit for your research. If you are not sure, read the appropriate sections before making your selection.
Life sciences Behavioural & social sciences  Ecological, evolutionary & environmental sciences
For a reference copy of the document with all sections, see nature.com/documents/nr-reporting-summary-flat.pdf
Life sciences study design
All studies must disclose on these points even when the disclosure is negative.
Sample size The sample size for this interim report was not based on statistical hypothesis testing. A total of 45 participants were enrolled in this part of 
the study. For the purposes of tolerability and dose escalation study a total of 15 participants (12 receiving vaccine and 3 receiving placebo) was deemed sufficient for a dosing finding phase study. 
Data exclusions All safety and immunogenicity data that were available at the time of the data snapshot were included in the interim report. No data were excluded from the analyses. 
Replication This is an interim report of an ongoing human clinical trial. There was no attempt at replication of study findings
Randomization This is an randomized controlled trial. Study participants were randomly assigned to a vaccine group using an interactive web based response technology system with each group comprising 15 participants (12 active vaccine recipients and 3 placebo recipients). 
Blinding This is an observer blinded study which is investigator blinded but Sponsor unblinded during Stage 1 (the stage from which data in the manuscript are presented). Investigators were unblinded to group level data but not subject level data for the purposes of interpretation and summary of the results included in this interim report. 
Reporting for specific materials, systems and methods
We require information from authors about some types of materials, experimental systems and methods used in many studies. Here, indicate whether each material, system or method listed is relevant to your study. If you are not sure if a list item applies to your research, read the appropriate section before selecting a response. 
Materials & experimental systems
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Antibodies
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ChIP seq
Flow cytometry
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Human research participants
Policy information about studies involving human research participants
Population characteristics Study participants were healthy men or women 18 55 years of age. Key exclusion criteria included individuals with known infection with human immunodeficiency virus, hepatitis C virus, or hepatitis B virus; immunocompromised individuals and those with a history of autoimmune disease; those with increased risk for severe COVID 19; previous clinical or microbiological diagnosis of COVID 19; receipt of medications intended to prevent COVID 19; previous vaccination with any coronavirus vaccine; a positive serological test for SARS CoV 2 IgM and/or IgG at the screening visit; and a SARS CoV 2 NAATpositive nasal swab within 24 hours before study vaccination.
Recruitment Study participants were recruited at the two individual sites and recruitment strategies were at the discretion of individual sites and could include identification of interested individuals from the sites local database or through advertising in the local community.  Once recruited participants were screened for eligibility based on pre specified protocol criteria. Eligible participants were then randomized to vaccine or placebo in a blinded manner. These processes therefore did not led themselves to enrollment biases however participants who did not know about the study may have had less of an opportunity to participate. 
Ethics oversight The study protocol was approved by the western institutional review board for one site and by the Langone Health New York University Institutional IRB prior to enrollment of any participants
Note that full information on the approval of the study protocol must also be provided in the manuscript.
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3 nature research  |  reporting summary April 2020Clinical data
Policy information about clinical studies
All manuscripts should comply with the ICMJE guidelines for publication of clinical research  and a completed CONSORT checklist must be included with all submissions.
Clinical trial registration ClinicalTrials.gov identifier: NCT04368728
Study protocol Details of protocol elements can be accessed from clinicaltrials.gov 
Data collection Data were collected at screening (up to 14 days before vaccination) and for randomized participants at the investigative site at 
baseline, 1 day, 7 days and 21 days,  after Dose 1, 7 days after dose 2 and up to 14 days after dose 2. Both safety and/or  serum collection for immunogenicity assessments were collected  for all stated time points. In addition, reactogenicity data were assessed through participant self reports via an electronic diary for 7 days after dose 1.  
Outcomes In this interim report, the following study primary endpoints are presented: the proportion of participants reporting prompted local reactions, systemic events, and use of antipyretic and/or pain medication within 7 days after vaccination, AEs and serious adverse events (SAEs) (available through up to ~45 days after Dose 1), and the proportion of participants with clinical laboratory abnormalities 1 and 7 days after vaccination and grading shifts in laboratory assessments between baseline and 1 and 7 days after Dose 1 and between Dose 2 and 7 days after Dose 2.  Secondary endpoints included: SARS CoV 2 neutralizing geometric mean titers (GMTs); SARS CoV 2 RBD binding IgG geometric mean concentrations (GMCs) 7 and 21 days after Dose 1 and 7 and 14 days after Dose 2
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