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Pi Is 1473309920308434
Pi Is 1473309920308434
Summary
Background With the unprecedented morbidity and mortality associated with the COVID-19 pandemic, a vaccine Lancet Infect Dis 2020
against COVID-19 is urgently needed. We investigated CoronaVac (Sinovac Life Sciences, Beijing, China), an Published Online
inactivated vaccine candidate against COVID-19, containing inactivated severe acute respiratory syndrome November 17, 2020
https://doi.org/10.1016/
coronavirus 2 (SARS-CoV-2), for its safety, tolerability and immunogenicity. S1473-3099(20)30843-4
See Online/Comment
Methods In this randomised, double-blind, placebo-controlled, phase 1/2 clinical trial, healthy adults aged https://doi.org/10.1016/
18–59 years were recruited from the community in Suining County of Jiangsu province, China. Adults with SARS- S1473-3099(20)30870-7
CoV-2 exposure or infection history, with axillary temperature above 37·0°C, or an allergic reaction to any vaccine For the Chinese translation
component were excluded. The experimental vaccine for the phase 1 trial was manufactured using a cell factory of the abstract see Online
for appendix 1
process (CellSTACK Cell Culture Chamber 10, Corning, Wujiang, China) , whereas those for the phase 2 trial were
produced through a bioreactor process (ReadyToProcess WAVE 25, GE, Umea, Sweden) . The phase 1 trial was *Joint first authors
done in a dose-escalating manner. At screening, participants were initially separated (1:1), with no specific †Contributed equally
randomisation, into two vaccination schedule cohorts, the days 0 and 14 vaccination cohort and the days 0 and 28 Department of Microbiology,
Zhejiang Provincial Center
vaccination cohort, and within each cohort the first 36 participants were assigned to block 1 (low dose CoronaVac for Disease Control and
[3 μg per 0·5 mL of aluminium hydroxide diluent per dose) then another 36 were assigned to block 2 (high-dose Prevention, Hangzhou, China
Coronavc [6 μg per 0·5 mL of aluminium hydroxide diluent per dse]). Within each block, participants were randomly (Prof Y Zhang PhD);
assigned (2:1), using block randomisation with a block size of six, to either two doses of CoronaVac or two doses of Sinovac Biotech, Beijing,
China (G Zeng PhD, Ya Hu MSc,
placebo. In the phase 2 trial, at screening, participants were initially separated (1:1), with no specific randomisation, W Han MSc, W Yin, MBA,
into the days 0 and 14 vaccination cohort and the days 0 and 28 vaccination cohort, and participants were randomly Yu Hu MPH); Jiangsu Provincial
assigned (2:2:1), using block randomisation with a block size of five, to receive two doses of either low-dose Center for Disease Control and
CoronaVac, high-dose CoronaVac, or placebo. Participants, investigators, and laboratory staff were masked to Prevention, Nanjing, China
(H Pan MSc, K Chu MSc,
treatment allocation. The primary safety endpoint was adverse reactions within 28 days after injection in all R Tang BA, Prof J Li PhD,
participants who were given at least one dose of study drug (safety population). The primary immunogenic outcome Prof F Zhu MD); National
was seroconversion rates of neutralising antibodies to live SARS-CoV-2 at day 14 after the last dose in the days 0 and Institutes for Food and Drug
14 cohort, and at day 28 after the last dose in the days 0 and 28 cohort in participants who completed their Control, Beijing, China
(Prof C Li PhD, Z Chen MSc);
allocated two-dose vaccination schedule (per-protocol population). This trial is registered with ClinicalTrials.gov, Suining County Center for
NCT04352608, and is closed to accrual. Disease Control and Prevention,
Suining, Jiangsu Province, China
Findings Between April 16 and April 25, 2020, 144 participants were enrolled in the phase 1 trial, and between May 3 and (X Chen BA, X Liu BA, C Jiang BA,
Y Song BA); CAS Key Laboratory
May 5, 2020, 600 participants were enrolled in the phase 2 trial. 743 participants received at least one dose of of Infection and Immunity,
investigational product (n=143 for phase 1 and n=600 for phase 2; safety population). In the phase 1 trial, the National Laboratory of
incidence of adverse reactions for the days 0 and 14 cohort was seven (29%) of 24 participants in the 3 ug group, Macromolecules, Institute of
Biophysics, Chinese Academy of
nine (38%) of 24 in the 6 μg group, and two (8%) of 24 in the placebo group, and for the days 0 and 28 cohort was
Sciences, Beijing, China
three (13%) of 24 in the 3 μg group, four (17%) of 24 in the 6 μg group, and three (13%) of 23 in the placebo group. (M Yang PhD, Prof X Wang PhD)
The seroconversion of neutralising antibodies on day 14 after the days 0 and 14 vaccination schedule was seen in and Sinovac Life Sciences,
11 (46%) of 24 participants in the 3 μg group, 12 (50%) of 24 in the 6 μg group, and none (0%) of 24 in the placebo Beijing, China (Q Gao MSc)
group; whereas at day 28 after the days 0 and 28 vaccination schedule, seroconversion was seen in 20 (83%) of 24 in Correspondence to:
the 3 μg group, 19 (79%) of 24 in the 6 μg group, and one (4%) of 24 in the placebo group. In the phase 2 trial, the Dr Qiang Gao, Sinovac Life
Sciences, Beijing 100085, China
incidence of adverse reactions for the days 0 and 14 cohort was 40 (33%) of 120 participants in the 3 μg group, gaoq@sinovac.com
42 (35%) of 120 in the 6 μg group, and 13 (22%) of 60 in the placebo group, and for the days 0 and 28 cohort was or
23 (19%) of 120 in the 3 μg group, 23 (19%) of 120 in the 6 μg group, and 11 (18%) of 60 for the placebo group.
Dr Fengcai Zhu, Jiangsu
Seroconversion of neutralising antibodies was seen for 109 (92%) of 118 participants in the 3 μg group, 117 (98%) Provincial Center for Disease
of 119 in the 6 μg group, and two (3%) of 60 in the placebo group at day 14 after the days 0 and 14 schedule; whereas Control and Prevention, Nanjing
at day 28 after the days 0 and 28 schedule, seroconversion was seen in 114 (97%) of 117 in the 3 μg group, 118 (100%) 210009, China
jszfc@vip.sina.com
of 118 in the 6 μg group, and none (0%) of 59 in the placebo group.
Interpretation Taking safety, immunogenicity, and production capacity into account, the 3 μg dose of CoronaVac is the
suggested dose for efficacy assessment in future phase 3 trials.
Funding Chinese National Key Research and Development Program and Beijing Science and Technology Program.
Research in context
Evidence before this study 97·6% at 14 days after a day 0 and 21 vaccination schedule.
We searched PubMed and the American Medical Association The clinical study of CoronaVac can further provide safety and
website on Aug 13, 2020, for published research articles, with immunogenic evidence for the inactivated vaccine.
no language or date restrictions, using the search terms of
Added value of this study
“SARS-CoV-2”, “COVID-19”, “vaccine”, and “clinical trial”.
In this first in-human study of CoronaVac, we used a phase 1/2
The search results showed that the COVID-19 pandemic
study design to screen the safety of two doses and
resulted in an unprecedented race to develop an effective
two vaccination schedules in a dose-escalation study in a small
vaccine. We identified preclinical data on three immunisations
cohort before expanding the study to a larger cohort to explore
using two different doses of CoronaVac (3 μg and 6 μg per
the immunogenicity of the vaccine in healthy adults.
dose), an inactivated whole virus vaccine against COVID-19
The immune response in the phase 2 study was substantially
developed by Sinovac Life Sciences (Beijing, China),
higher than in the phase 1 study, which might be due to the
providing partial or complete protection in macaques against
difference in preparation process of vaccine batches used in
SARS-CoV-2 challenge, without observable antibody-
phase 1 and 2 resulting in a higher proportion of intact
dependent enhancement of infection. We also identified a
spike protein on the purified inactivated SARS-CoV-2 virions in
phase 2 clinical trial of another inactivated vaccine developed
the vaccine used in phase 2 than that used in phase 1.
by Sinopharm (Beijing, China), which showed the incidence of
adverse reactions was 19·0% within 28 days after two doses of Implications of all the available evidence
vaccine (5 μg in 0·5 mL of diluent) in a day 0 and 21 vaccination Data from this study support the approval of emergency use of
schedule, and the seroconversion rates of the neutralising CoronaVac in China, and three phase 3 clinical trials that are
antibody detected by plaque reduction neutralisation test was ongoing in Brazil, Indonesia, and Turkey.
(1:1), with no specific randomisation, to one of two dose of vaccine, and only after a successful safety
vaccination schedules, with either a 14-day interval (the observation 7 days after the first dose was the trial able to
day 0 and 14 vaccination cohort) or a 28-day interval (the proceed and participants in block 2 be given the high
day 0 and 28 vaccination cohort) between doses. Within dose of vaccine. The criteria that had to be met from the
each cohort, the first 36 participants (block 1) were 7-day safety observation were that no life-threatening
randomly assigned to either the low dose vaccine or adverse events occur, no more than 15% of vaccinated
placebo, and then after 7 days of follow-up for safety after participants report severe adverse events, and no other
the first dose, another 36 (block 2) were randomly assigned safety concerns in the opinion of the data monitoring
to either high-dose vaccine or placebo. Phase 2 was committee (DMC) occur. The same conditions needed to
initiated after all participants in phase 1 has finished a be met 7 days after the first dose in block 2 of the phase 1
7-day safety observation period after the first dose. As in trial before the study could proceed to the phase 2 trial.
phase 1, participants were recruited and allocated (1:1) CoronaVac is an inactivated vaccine candidate against
with no specific randomisation to one of the two COVID-19, created from African green monkey kidney
vaccination-schedule cohorts, and then randomly assigned cells (Vero cells) that have been inoculated with SARS-
within each cohort to either low-dose vaccine, high-dose CoV-2 (CN02 strain). At the end of the incubation period,
vaccine, or placebo. the virus was harvested, inactivated with β-propiolactone,
Participants were eligible if they were healthy and aged concentrated, purified, and finally absorbed onto alu
18–59 years. The key exclusion criteria were high-risk minium hydroxide. The aluminium hydroxide complex
epidemiology history within 14 days before enrolment was then diluted in a sodium chloride, phosphate-
(eg, travel or residence history in Wuhan city and buffered saline, and water solution before being sterilised
surrounding areas or other communities with case reports; and filtered ready for injection. The placebo is just the
contact history with someone infected with SARS-CoV-2); aluminium hydroxide diluent solution with no virus.
SARS-CoV-2 specific IgG or IgM positive in serum; Both the vaccine and placebo were prepared in a Good
positive PCR test for SARS-CoV-2 from a pharyngeal or Manufacturing Practice-accredited facility of Sinovac Life
anal swab sample; axillary temperature of more than Sciences (Beijing, China) that is periodically inspected by
37·0°C; and known allergy to any vaccine component. A the Chinese National Medical Products Administration
complete list of exclusion criteria is in the protocol. committee for compliance. Vaccine of 3 μg and 6 μg in For the protocol see http://www.
Written informed consent was obtained from each 0·5 mL of aluminium hydroxide diluent per dose jscdc.cn/jkfw/kygz/202009/
t20200930_69600.html
participant before enrolment. The clinical trial protocol and placebo in ready-to-use syringes were administered
and informed consent form were approved by the intramuscularly according to the dosing schedule of
Jiangsu Ethics Committee (JSJK2020-A021–02). This either day 0 and day 14, or day 0 and day 28, depending
study was conducted in accordance with the requirements on the cohort. These vaccine doses had been found to be
of Good Clinical Practice of China and the International sufficient for protection against SARS-CoV-2 challenge in
Conference on Harmonisation. macaques.15 Cultivation technology by cell factory system
(CellSTACK Cell Culture Chamber 10, Corning, Wujiang,
Randomisation and masking China) was used in the preparation of the vaccine used in
In both phase 1 and 2, no specific randomisation was the phase 1 trial. However, for the phase 2 trial, we used a
used when allocating participants to the vaccinations highly automated bioreactor (ReadyToProcess WAVE 25,
schedule cohorts. In phase 1, participants in blocks 1 and GE, Umea, Sweden) to produce the vaccine to increase
2 in each schedule cohort were randomly assigned (2:1) to vaccine production capacity. After the immunogenicity
either CoronaVac or placebo, and in phase 2, participants results of the trial were obtained, we discovered that the
in each schedule cohort were randomly assigned (2:2:1) to change in manufacture of the vaccine optimised the cell
either low-dose CoronaVac, high-dose CoronaVac, or culture and resulted in higher intact spike protein
placebo. The randomisation codes for each vaccination content of the vaccine batch for the phase 2 trial, which
schedule cohort were generated individually, using block was unexpected. However, we were not aware of this
randomisation with a block size of six in phase 1 and a antigen-level difference between the vaccine batches for
block size of five in phase 2, using SAS software (version the phase 1 and 2 trials when we obtained the ethical
9.4). The randomisation code was assigned to each approval for the trials.
participant in sequence in the order of enrolment, and For the first 7 days after each dose, participants were
then the participants received the investigational products required to record the injection-site adverse events
labelled with the same code. The vaccine and the placebo (eg, pain, redness, swelling), or systemic adverse events
are identical in appearance. All participants, investigators, (eg, allergic reaction, cough, fever) on paper diary cards.
and laboratory staff were masked to treatment allocation. From day 8 to day 28 after each dose (and day 8 to day 14
for the first dose of the days 0 and 14 vaccination cohort),
Procedures safety data were collected by spontaneous report from
The phase 1 clinical trial was run in a dose-escalation the participants combined with the regular visit (which
manner. First, participants in block 1 were given the low occurred on day 8 and day 28 after each dose, and on day
8 and day 14 for the first dose in the days 0 and 14 Outcomes
vaccination schedule cohort). Serious adverse events The primary safety endpoint was any adverse reactions
were collected through the trial and will be collected until within 28 days after each dose of study drug. Secondary
6 months after the last dose. The reported adverse events safety endpoints were any abnormal changes in labora
were graded according to the China National Medical tory measurements at day 3 and in serum inflammatory
Products Administration guidelines.16 The causal factors 7 days after each dose of study drug. The secondary
association between adverse events and vaccination was safety endpoints were prespecified only in the phase 1 trial.
determined by the investigators. The primary immunogenic endpoint was the sero
In the phase 1 trial, blood and urine samples were conversion of neutralising antibodies to live SARS-CoV-2
taken on day 3 after each dose and tested to investigate at day 14 after the last dose in the days 0 and 14 vaccination
any abnormal changes of the haematology and bio cohort, or day 28 after the last dose in the days 0 and 28
chemistry indexes. 7 days after each dose, blood vaccination cohort. Secondary immunogenic endpoints
and urine samples were taken to measure serum infla were geometric mean titres (GMTs) of neutralising
mmatory factors including IL-2, IL-6, and TNF-α using antibodies to live SARS-CoV-2, RBD-specific IgG,
the solid phase sandwich ELISA method to explore the S-specific IgG, and IgM. Exploratory endpoints were
underlying pathological immune responses. Blood T-cell responses and, post hoc, GMTs of neutralising
samples were collected at days 0 (baseline), 7, 14, 21, 28, antibodies to psuedovirus. Seroconversion of antibodies
and 42 from participants in the day 0 and 14 vaccination was defined as a change from seronegative at baseline to
cohort, and days 0, 28, 35, 42, and 56 from participants in seropositive or a four-fold titre increase if the participant
the days 0 and 28 vaccination schedule cohort, to was seropositive at baseline. The positive cutoff of
determine the levels of neutralising antibodies, receptor- the neutralising antibodies to live SARS-CoV-2 was 1/8,
binding domain (RBD)-specific IgG, S-specific IgG, and neutralising antibodies to pseudovirus was 1/30, and
IgM. Additionally, T-cell responses were determined via RBD-specific IgG was 1/160. Regarding the ELISpot
IFN-γ detection on day 14 after each dose. measured T-cell response, the results were expressed as
In the phase 2 trial, blood samples were collected on the number of spot-forming cells (SFCs) per 100 000 cells.
day 0, 28, and 56 from participants in the days 0 Other secondary endpoints are listed in the appendix 2
and 14 cohort, and on day 56 from participants in the (p 4), including 6 month outcomes that are not available
days 0 and 28 cohort, to determine the levels of yet, which will be reported elsewhere.
neutralising antibodies and RBD-specific IgG.
The neutralising antibodies to live SARS-CoV-2 (virus Statistical analysis
strain SARS-CoV-2/human/CHN/CN1/2020, GenBank We assessed the safety endpoints in the safety
number MT407649.1) were quantified using a micro population, which included all participants who received
cytopathogenic effect assay17 with a minimum four-fold at least one dose of study drug. We assessed immuno
dilution, and neutralising antibodies to pseudovirus18 genic endpoints in the per-protocol population, which
were quantified with a minimum ten-fold dilution. included all participants who completed their assigned
The S-specific IgG and IgM were detected using the two-dose vaccination schedule and with available
chemiluminescence qualitative kit (Auto Biotechnology, antibody results.
Zhengzhou, China). These antibody detection tests We did not determine the sample size on the basis of a
were done by the National Institute for Food and Drug statistical power calculation, but followed the requirement
Control (Beijing, China). of the National Medical Products Administration in
Additionally, antibody titres for RBD-specific IgG China—ie, recruitment of at least of 20–30 participants in
were quantified using the in-house ELISA kit from phase 1 and 500 participants in phase 2.
Sinovac, with a minimum 160-fold dilution. T-cell We used the Pearson χ² test or Fisher’s exact test for the
response was determined with the ELISpot method analysis of categorical outcomes. We calculated 95% CIs
using a commercial kit (Human IFN γ ELISpotPRO for all categorical outcomes using the Clopper-Pearson
[3420-2AST-10, AID]; Mabtech, Stockholm, Sweden). method. We calculated GMTs and corresponding
Further information on all methods is in the 95% CIs on the basis of standard normal distribution of
See Online for appendix 2 appendix 2 (pp 1–3). Additionally, in a post-hoc analysis, the log-transformed antibody titre. We used the ANOVA
we tested serum samples from 117 convalescent patients method to compare the log-transformed antibody titre.
who had previously had COVID-19 collected in the When the comparison among all three groups showed
hospitals for neutralising antibodies to live SARS-CoV-2 significant difference, we then did pairwise comparisons.
using the same method as for the detection of serum Hypothesis testing was two-sided and we considered
neutralising antibodies to live SARS-CoV-2 in the p values of less than 0·05 to be significant.
phase 1 and 2 trials, to give a comparison of the vaccine- An independent data monitoring committee con
induced and infection-induced humoral immunity. sisted of one independent statistician, one clinician,
Written informed consent was obtained from all these and one epidemiologist was established before com
convalescent patients. mencement of the study. Safety data were assessed and
reviewed by the committee to ensure the suspension 662 individuals were screened and 600 participants were
criteria of the dose-escalation part of phase 1 were not enrolled in the phase 2 trial. 743 participants received at
met and allow the further proceeding of the clinical trial. least one dose of the investigational product (143 for
We used SAS (version 9.3) for all analyses. This trial is phase 1 and 600 for phase 2) and were included in the
registered with ClinicalTrials.gov, NCT04352608. safety population (figure 1). 143 participants in phase 1
and 591 participants in phase 2 were eligible for the
Role of the funding source immunogenic evaluation (per-protocol population;
The funder of the study had no role in study design, figure 1). Baseline demographic characteristics of the
data collection, data analysis, data interpretation, or participants in the safety population at enrolment were
writing of the report. All the authors have full access to similar among the treatment groups in terms of sex,
all the data in the study and the corresponding authors nationality, and mean age (table 1).
had final responsibility for the decision to submit for In the phase 1 trial, the overall incidence of adverse
publication. reactions was seven (29%) of 24 participants in the 3 μg
group, nine (38%) of 24 in the 6 μg group, and two (8%)
Results of 24 in the placebo group in the days 0 and 14 vaccination
Between April 16 and April 25, 2020, 185 individuals cohort; and three (13%) of 24 in the 3 μg group,
were screened and 144 participants were enrolled in the four (17%) of 24 in the 6 μg group, and three (13%) of 23
phase 1 trial, and between May 3 and May 5, 2020, in the placebo group in the days 0 and 28 vaccination
A Phase 1: days 0 and 14 vaccination cohort B Phase 1: days 0 and 28 vaccination cohort
21 excluded 20 excluded
20 not eligible 19 not eligible
1 withdrew 1 withdrew
72 enrolled 72 enrolled
1 with-
drew
24 given first dose* 12 given first dose* 24 given first dose† 12 given first dose† 24 given first dose* 12 given first dose* 24 given first dose† 11 given first dose†
24 given second 12 given second 24 given second 12 given second 24 given second 12 given second 24 given second 11 given second
dose dose dose dose dose dose dose dose
C Phase 2: days 0 and 14 vaccination cohort D Phase 2: days 0 and 28 vaccination cohort
33 excluded 29 excluded
11 not eligible 9 not eligible
22 withdrew 20 withdrew
120 randomly 120 randomly 60 randomly 120 randomly 120 randomly 60 randomly
assigned to assigned to assigned to assigned to assigned to assigned to
6 μg group 3 μg group placebo group 6 μg group 3 μg group placebo group
120 given first dose 120 given first dose 60 given first dose 120 given first dose 120 given first dose 60 given first dose
119 given second 120 given second 60 given second 118 given second 117 given second 60 given second
dose dose dose dose dose dose
1 given placebo‡
120 included in 120 included in 60 included in 120 included in 120 included in 60 included in
safety safety safety safety safety safety
population population population population population population
119 included in 118 included in 60 included in 118 included in 117 included in 59 included in
per-protocol per-protocol per-protocol per-protocol per-protocol per-protocol
population population§ population population population population¶
A Phase 1, days 0 and 14 vaccination cohort B Phase 1, days 0 and 28 vaccination cohort
50 3 μg group (n=24) 3 μg group (n=24)
45 6 μg group (n=24) 6 μg group (n=24)
40 Placebo group (n=24) Placebo group (n=23)
35
Incidence (%)
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50 3 μg group (n=120) 3 μg group (n=120)
45 6 μg group n=120) 6 μg group (n=120)
40 Placebo group (n=60) Placebo group (n=60)
35 * *
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30
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Figure 2: Incidence of adverse reactions reported within 28 days after second dose of study drug, in the days 0 and 14 vaccination cohort in phase 1 (A) and phase 2 (C) and in the days 0
and 28 vaccination cohort in phase 1 (B) and phase 2 (D)
Adverse reactions refer to the adverse events related to the vaccination. Rare injection-site symptoms reported only in the days 0 and 14 vaccination cohort are not shown in the figure and are listed in
appendix 2 along with all adverse reactions after the first and second dose (pp 4–13). *The p value of comparison among three groups is significant for the incidence of any injection-site symptoms
(p=0·02) and injection-site pain (p=0·04).
Additionally, ten (7%) of 143 participants in phase 1 had a 6 μg group (25·9 [14·6–46·1) versus none of 23 in the
clinically significant increase of laboratory indicators at placebo group (2·0 [2·0–2·0]) at 14 days after the second
day 3 after vaccination (appendix 2 pp 15–16), but none dose, and 20 (83%) in the 3 μg group (19·0 [13·2–27·4]
was considered to be related to the vaccination. No versus 19 (79%) in the 6 μg group (29·6 [17·9–48·9])
significant increases in inflammatory factors in serum versus one (4%) in the placebo group (2·2 [1·8–2·8]) at
were detected at day 7 after each dose (appendix 2 pp 17–18). 28 days after the second dose in the days 0 and 28
At baseline, none of the participants in the phase vaccination cohort (table 2, figure 3; appendix 2 p 19).
1 trial had any detectable neutralising antibodies to live The seroconversion rates of RBD-specific IgG were 20
SARS-CoV-2. The seroconversion rates of neutralising (83%) of 24 participants in the 3 μg group (GMT 465·8
antibodies were 11 (46%) of 24 participants in the 3 μg [95% CI 277·6–781·7] versus 24 (100%) of 24 participants
group (GMT 5·6 [95% CI 3·6–8·7]) versus 12 (50%) of in the 6 μg group (987·0 [647·8–1504·0]) versus two (8%)
24 participants in the 6 μg group (7·7 [5·2–11·5]) of 24 participants in the placebo group (84·8 [78·0–92·1])
versus none of 24 participants in the placebo group at 14 days after the second dose, and 21 (88%) in the 3 μg
(2·0 [2·0–2·0]) at 14 days after the second dose, and group (465·8 [288·1–753·1]) versus 24 (100%) in the
six (25%) participants in the 3 μg group (5·4 [3·6–8·1] 6 μg group (1395·9 [955·2–2039·7]) versus two (8%) in
versus 20 (83%) in the 6 μg group (15·2 [11·2–20·7]) the placebo group (89·8 [76·1–105·9]) at 28 days after
versus none in the placebo group (2·0 [2·0–2·0]) the second dose in the days 0 and 14 vaccination
at 28 days after the second dose in the days 0 and 14 cohort; and 24 (100%) of 24 participants in the 3 μg
vaccination cohort; and 19 (79%) of 24 participants in the group (1365·1 [881·4–2086·4]) versus 24 (100%) of 24
3 μg group (16·0 [10·4–24·7]) versus 20 (83%) of 24 in the participants in the 6 μg group (2152·7 [1446·1–3204·6])
A Phase 1, days 0 and 14 vaccination cohort B Phase 1, days 0 and 28 vaccination cohort C Phase 2, days 0 and 14 vaccination cohort D Phase 2, days 0 and 28 vaccination cohort
2048 3 μg group p=0·027 p=0·0006
Reciprocal antibody titre of neutralising
1024 6 μg group
Placebo group 34·5 30·1
antibody to live SARS-CoV-2
E Phase 1, days 0 and 14 vaccination cohort F Phase 1, days 0 and 28 vaccination cohort G Phase 2, days 0 and 14 vaccination cohort H Phase 2, days 0 and 28 vaccination cohort
p=0·024
p=0·024 p=0·0006 p=0·018 p=0·013
1365·4 1318·2
Reciprocal antibody titre of RBD-specific IgG
Days after second dose Days after second dose Days after second dose Days after second dose
Figure 3: Antibody titres of neutralising antibodies to live SARS-CoV-2 (A–D) and RBD-specific IgG (E–H) induced after two doses of CoronaVac or placebo given in the days 0 and 14 and
days 0 and 28 vaccination cohorts, in the phase 1 and phase 2 trials
The error bars indicate the 95% CI of the GMT and the spots indicated the individual antibody titres, with the numbers above the spots showing the GMT estimate. Only p values for significant
differences are shown on the figure, all p values for all data are in appendix 2 (p 19). GMT=geometric mean titre. RBD=receptor binding domain. SARS-CoV-2=severe acute respiratory syndrome
coronavirus 2.
(GMT 163·7 [95% CI 128·5–208·6]; table 2, figure 3; pseudovirus was 0·82 (0·76–0·88) using the antibody
appendix 2 p 24). The seroconversion rates of RBD-specific titre at 14 days after the second dose (no data taken at
IgG were 111 (97%) of 115 participants in the 3 μg group day 28). The correlation coefficient between the
(GMT 1094·3 [95% CI 936·7–1278·4]) versus 118 (100%) of neutralising antibody to pseudovirus and RBD-specific
118 participants in the 6 μg group (1365·4 [1160·4–1606·7]) IgG was 0·73 (0·66–0·80) using the antibody titre at 14
versus none of 56 participants in the placebo group days after the second dose (no data taken at day 28;
(81·0 [79·0–83·0]) at 14 days after the second dose and appendix 2 p 24).
111 (97%) of 114 in the 3 μg group (1053·7 [911·7–1217·7])
versus 118 (100%) of 118 in the 6 μg group Discussion
(1318·2 [1156·9–1501·9]) versus none of 57 in the placebo We found that two doses of CoronaVac at different concen
group (80·0 [80·0–80·0]) at 28 days after the second dose trations and using different dosing schedules were well
in the day 0 and 14 vaccination cohort; and 116 (99%) of tolerated and moderately immunogenic in healthy adults
117 in the 3 μg group (1783·6 [1519·3–2093·8]) versus aged 18–59 years. The incidence of adverse reactions in the
117 (100%) of 117 in the 6 μg group (2287·5 [2038·2–2567·3]) 3 μg and 6 μg group were similar, indicating no dose-
versus four (7%) of 59 in the placebo group related safety concerns but more long-term follow-up is
(87·9 [79·7–96·9]) at 28 days after the second dose in the needed. Furthermore, most adverse reactions were mild,
days 0 and 28 vaccination cohort (table 2, figure 3). with the most common symptom being injection-site pain,
Based on the pooled data of the phase 1 and 2 trials which is in accordance with previous findings for another
(two vaccination cohorts pooled), the correlation co inactivated COVID-19 vaccine from Sinopharm (Beijing
efficient between the neutralising antibody to live SARS- China).14 Compared with other COVID-19 vaccine candi
CoV-2 and RBD-specific IgG was 0·85 (95% CI dates, such as viral-vectored vaccines or DNA or RNA
0·82–0·92) using the antibody titre at 28 days after the vaccines, the occurrence of fever after vaccination with
second dose of vaccine, and was 0·80 (0·75–0·86) using CoronaVac was relatively low.10,11,13
the titre 14 days after the second. The correlation Over the course of the phase 1/2 trial, we changed the
coefficient between the neutralising antibody to production process of the vaccine from the use of a cell
live SARS-CoV-2 and the neutralising antibody to factory process (which was used in our preclinical and
phase 1 study to generate a 50 L culture of Vero cells) to that CoronaVac could provide satisfying protection
use of a bioreactor for phase 2. The bioreactor process against COVID-19 on the basis of the following three
enabled use to optimise the process for growing cells, reasons. First, from the experiences of other vaccines,
with precise control over cell culture parameters like such as the enterovirus 71 and varicella vaccines, most of
dissolved oxygen, pH, and carbon dioxide and oxygen the surrogate endpoints based on neutralising antibody
gas levels. We made this change to increase vaccine titres have ranged from 8 to 24.20,21 Second, our preclinical
production capacity and meet biosafety requirements. study15 indicated that the neutralising antibody titres of
Pre-clinical data for each phase trial (data not shown) 1/24 elicited in macaque models conferred complete
indicated that the safety profiles of vaccines prepared via protection against SARS-CoV-2. Third, although several
the new bioreactor process and old process are similar. studies have found that antibody responses generated
Notably, immune responses in phase 2 were much better from natural infection with corona viruses (eg, SARS-
than those recorded in phase 1, with seroconversion rates CoV-2, severe acute respiratory syndrome coronavirus,
over 90% in both the 3 μg and 6 μg groups. To investigate and Middle East respi ratory syndrome coronavirus)
the reason for this change, we did a protein composition might decrease substantially over time,22–24 reinfection in
analysis of the purified inactivated SARS-CoV-2 virions these patients has rarely been reported,25–27 which indicates
and found that the bioreactor-produced vaccine had a that immunological memory might have an important
higher redundancy of intact spike protein (molecular role of prevention of re-infections. Therefore, the antibody
mass approximately 180 kDa) than did the vaccine level itself might not be the key for a successful COVID-19
produced via the cell factory process (appendix 2 p 27). vaccine, but rather the establishment of a recallable
Quantitative analysis showed that the intact spike protein specific immune response to SARS-CoV-2. Furthermore,
accounted for approximately 3·7% of total protein mass the efficacy of the investigational vaccine and its surrogate
of the vaccine used in phase 1 and approximately 7·0% of endpoint need to be determined in a future phase 3 trial.
total protein mass of the vaccine used in phase 2 trials. Additionally, comparability of our serum antibody results
Electron microscopic examination of the samples further with those of other COVID-19 vaccine studies is restricted.
verified that the average number of spikes per virion of Two participants in the placebo group in the phase 1
the viral sample used in the phase 2 trial was almost trial and four in the placebo group in the phase 2 trial
double the number of spikes per virion of the sample had seroconversion of anti-RBD IgG after vaccination,
used in phase 1 trial (appendix 2 p 27). These observations and one participant given placebo in the phase 1 trial and
highlight the importance of developing an optimum two in the phase 2 trial had seroconversion of neutralising
manu facturing process and the integration of multi antibodies after vaccination.
disciplinary techniques, such as genomics and structural CoronaVac was well tolerated and induced humoral
biology to support a new era of precision vaccinology. responses against SARS-CoV-2, which supported the
The immune response induced by 3 μg and 6 μg of approval of emergency use of CoronaVac in China, and
vaccine in 0·5 mL of diluent per dose was similar in this three phase 3 clinical trials that are ongoing in Brazil
study. As anticipated, after two doses of vaccine, immune (NCT04456595), Indonesia (NCT04508075), and Turkey
responses induced by the days 0 and 28 vaccination (NCT04582344). Taking safety, immunogenicity, and
schedule were larger than those induced by the days 0 production capacity into account, the low dose of 3 μg of
and 14 vaccination schedule, regardless of the dose. CoronaVac in 0·5 mL of diluent, with a day 0 and 14
However, quick antibody responses could be induced vaccination schedule, is being investigated in these
within a relatively short time by using a day 0 and 14 ongoing trials. And the days 0 and 28 vaccination
vaccination schedule, which might be suitable for schedule with 3 μg of Coronavac in 0·5 mL of diluent
emergency use and is of vital importance during will also be investigated in future phase 3 clinical trials.
the COVID-19 pandemic. Regarding the days 0 The protective efficacy of CoronaVac remains to be
and 28 vaccination schedule, a more robust antibody determined.
response was generated and longer persistence could be Our study had several limitations. First, we did not
expected than with the days 0 and 14 schedule, which assess the T cell responses in the phase 2 trial; however,
supports potential routine use of the vaccine according to the response of type 1 T-helper cells and type 2 T-helper
this schedule when the epidemic risk of COVID-19 is cells induced by CoronaVac will be studied in the ongoing
low. However, the actual immune persistence of the phase 3 study in Brazil (NCT04456595). Second, we only
two schedules needs to be verified in future studies. reported immune response data for healthy adults, and
In the phase 2 trial, the level of neutralising antibodies did not include individuals from more susceptible
included by the vaccine at day 28 after the last dose of groups in our study population (eg, older individuals
vaccine ranged from a GMT of 23·8 to 65·4, depending [aged ≥60 years] or with comorbidities); and data on
on the vaccination schedule, which was lower than those immune persistence is not yet available, which need to
of convalescent patients who previously had COVID-19 be further studied. Third, the calculated p values
with an average GMT level of 163·7, tested by the same presented in this study cannot support any powerful
method in the same laboratory.19 However, we still think statistical conclusions, and are only for reference and so
23 Prévost J, Gasser R, Beaudoin-Bussières G, et al. Cross-sectional 26 Chan PKS, Lui G, Hachim A, et al. Serologic responses in healthy
evaluation of humoral responses against SARS-CoV-2 spike. adult with SARS-CoV-2 reinfection, Hong Kong, August 2020.
Cell Rep Med 2020; 1: 100126. Emerg Infect Dis 2020; published online Oct 22.
24 Payne DC, Iblan I, Rha B, et al. Persistence of antibodies against https://doi.org/10.3201/eid2612.203833.
Middle East respiratory syndrome coronavirus. Emerg Infect Dis 2016; 27 Huang AT, Garcia-Carreras B, Hitchings MDT, et al. A systematic
22: 1824–26. review of antibody mediated immunity to coronaviruses: kinetics,
25 Zhang K, Lau JY-N, Yang L, Ma Z-G. SARS-CoV-2 reinfection in two correlates of protection, and association with severity. Nat Commun
patients who have recovered from COVID-19. Precis Clin Med 2020; 2020; 11: 4704.
published online Sept 4. https://doi.org/10.1093/pcmedi/pbaa031.