22
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23
Figure S1 Preparation of RBD
-
mi3 nanoparticles
, related to Figure 1.
(A)
H
ypothesis
illustrating potential mechanism
for mosaic RBD
-
nanoparticle
induction
of cross
-
reactive
Abs
. Left:
Both Fabs of
a strain
-
specific
membrane
-
bound
BCR
can
bind to a strain
-
specific epitope (
pale yellow
triangle) on
yellow
antigens attached to a homotypic
nano
particle.
Middle:
Strain
-
specific
BCR
s
can only
bind with
one Fab
to
a
strain
-
specific epitope (triangle) on
yellow
antigen attached to a mosaic
nano
particle. Right:
Cross
-
reactive
BCR
s can bind with
both
Fabs
to
a
common epitope present on
adjacent
antigens
(green circle
s
)
attached to a mosaic
particle, but not to strain
-
specific epitopes (triangles).
(
B
)
Schematic of c
onstruction of mosaic
-
8b, homotypic SARS
-
2, and
a
dmix
-
8b RBD
-
mi3
nanoparticles
made
using models constructed with coordinates of an
RBD
(PDB 7BZ5),
SpyCatcher (PDB 4MLI), and
an
i3
-
01
nanoparticle
(PDB 7B3Y)
.
(
C
)
Superose 6 10/300
size exclusion chromatography
profile after RBD conjugations to mi3
showing peaks for RBD
-
mi3 nanoparticles.
(
D
)
Coomassie
-
stained
SDS
-
PAGE
gel
of RBD
-
coupled nanoparticles,
unconjugated
RBDs, and
free SpyCatcher003
-
mi3 particles (SC3
-
mi3).
.
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available under a
(which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made
The copyright holder for this preprint
this version posted February 9, 2024.
;
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doi:
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24
Table S1
Summary of vaccines and
immunogens
, r
elated to Figure 1
.
Animal Studies
Vacinated with
Vaccine Source
Immunized with
Immunogen Source
Figures
mosaic-8b
Caltech
homotypic SARS-2
Caltech
WA1/BA.1 rep-RNA
HDT
mosaic-8b
Caltech
admix-8b
Caltech
homotypic SARS-2
Caltech
Pfizer-like WA1
Helix Biotech
mosaic-8b
Caltech
mosaic-7
Caltech
Pfizer-like WA1 and WA1/BA.5 mRNA-LNP
Helix Biotech
mosaic-8b
Caltech
admix-8b
Caltech
homotypic SARS-2
Caltech
WA1 mRNA-LNP
Rockefeller
WA1 ChAdOx1
Jenner Institute, Oxford
mosaic-8b
Caltech
homotypic SARS-2
Caltech
UW, HDT, U Albany,
Creative Biosciences
2, S2
pre-vax BALB/c mice
3, S3
Helix Biotech
Pfizer-like WA1
Pfizer-like WA1 and WA1/BA.5 mRNA-LNP
WA1 ChAdOx1
WA1 mRNA-LNP
pre-vax NHPs
WA1 Spike, Bivalent (Beta/Delta) SHARP, Trivalent
(WA1/Beta/Delta) SHARP
Delivered via: repRNA-LION, repRNA gene gun, or DNA
pre-vax BALB/c mice
4, 5, S4
pre-vax Tg mice
Jenner Institute, Oxford
UPenn, Acuitas
6, S6, S7
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27
Figure S
2
Mosaic
-
8b immunizations in previously
-
vaccinated NHPs elicits cross
-
reactive
Ab responses
,
related to
Figure 2.
Data for ELISA and neutralization analyses for serum samples from weeks 4 and 12. Data for samples
from weeks 0, 2, 8, 10, and 22 are shown in Figure 2.
The mean of mean titers in panels C and E is
compared across immunizations by Tukey’s multiple comparison test with the Geisser
-
Greenhouse
correction (as calculated by GraphPad Prism), with pairing by viral strain.
Significant differences
between cohorts linked by horizontal lines are indicated by asterisks: p<0.05 = *, p<0.01 = **, p<0.001
= ***, p<0.0001
= ****.
(A
)
Left: Top: Stratification of pre
-
vaccinated NHPs into groups used for immunizations with
mosaic
-
8b, homotypic SARS
-
2, and WA1/BA.1 mRNA
.
Neutralization ID
50
values derived from
samples taken at week
-
8 in Figure 2A. Right: Numbers and color coding used to indicate
sarbecovirus strains within clades throughout the figure. (Bottom) Colors and symbols used to
indicate different immunizations (colors) and matched
versus mismatched viral strains (symbols).
(B) ELISA binding titers at the indicated weeks after priming with mosaic
-
8b, homotypic SARS
-
2,
or bivalent WA1/BA.1 mRNA
-
LNP represented as mean ED
50
values for serum IgG binding to
RBD or spike proteins from the indicated sarbecovirus strains (numbers and color coding as in
panel A).
.
CC-BY-NC 4.0 International license
available under a
(which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made
The copyright holder for this preprint
this version posted February 9, 2024.
;
https://doi.org/10.1101/2024.02.08.576722
doi:
bioRxiv preprint