1. Immunology and Inflammation
  2. Microbiology and Infectious Disease
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Escape from neutralizing antibodies by SARS-CoV-2 spike protein variants

  1. Yiska Weisblum
  2. Fabian Schmidt
  3. Fengwen Zhang
  4. Justin DaSilva
  5. Daniel Poston
  6. Julio CC Lorenzi
  7. Frauke Muecksch
  8. Magdalena Rutkowska
  9. Hans-Heinrich Hoffmann
  10. Eleftherios Michailidis
  11. Christian Gaebler
  12. Marianna Agudelo
  13. Alice Cho
  14. Zijun Wang
  15. Anna Gazumyan
  16. Melissa Cipolla
  17. Larry Luchsinger
  18. Christopher D Hillyer
  19. Marina Caskey
  20. Davide F Robbiani
  21. Charles M Rice
  22. Michel C Nussenzweig
  23. Theodora Hatziioannou  Is a corresponding author
  24. Paul D Bieniasz  Is a corresponding author
  1. Laboratory of Retrovirology, The Rockefeller University, United States
  2. Laboratory of Molecular Immunology The Rockefeller University, United States
  3. Laboratory of Virology and Infectious Disease The Rockefeller University, United States
  4. Lindsley F. Kimball Research Institute, New York Blood Center, United States
  5. Institute for Research in Biomedicine, Università della Svizzera italiana, Switzerland
  6. Howard Hughes Medical Institute, The Rockefeller University, United States
Research Article
Cite this article as: eLife 2020;9:e61312 doi: 10.7554/eLife.61312
9 figures, 3 tables and 1 additional file

Figures

Selection of SARS-CoV-2 S mutations that confer antibody resistance.

(A) Outline of serial passage experiments with replication-competent VSV derivatives encoding the SARS-CoV-2 S envelope glycoprotein and a GFP reporter (rVSV/SARS-CoV-2/GFP) in 293T/ACE2(B) cells in the presence of neutralizing antibodies or plasma. Each passage experiment was performed twice (once each with rVSV/SARS-CoV-2/GFP1D7 and rVSV/SARS-CoV-2/GFP2E1.) (B) Representative images of 293T/ACE2(B) cells infected with 1 × 106 PFU of rVSV/SARS-CoV-2/GFP in the presence or absence of 10 μg/ml of the monoclonal antibody C121. (C) Expanded view of the boxed areas showing individual plaques of putatively antibody-resistant viruses.

Figure 2 with 2 supplements
Analysis of S-encoding sequences following rVSV/SARS-CoV-2/GFP replication in the presence of neutralizing monoclonal antibodies.

(A–D) Graphs depict the S codon position (X-axis) and the frequency of non-synonymous substitutions (Y-axis) following the second passage (p2) of rVSV/SARS-CoV-2/GFP on 293T/ACE2(B) cells in the absence of antibody or plasma (A), or in the presence of 10 μg/ml C121 (B), C135 (C) or C144 (D). Results are shown for both rVSV/SARS-CoV-2/GFP variants (One replicate each for rVSV/SARS-CoV-2/GFP1D7 and rVSV/SARS-CoV-2/GFP2E1 - the frequency of 1D7 mutations is plotted as circles and 2E1 mutations as triangles).

Figure 2—figure supplement 1
Analysis of S-encoding sequences following rVSV/SARS-CoV-2/GFP replication in the presence of convalescent plasma COV-47 and COV-72.

(A–B) Graphs depict the S codon position (X-axis) and the frequency of non-synonymous substitutions (Y-axis) following the second, third or fourth passage (p2–p4) of rVSV/SARS-CoV-2/GFP on 293T/ACE2(B) cells in the presence of COV-47 plasma (A), or COV-72 plasma (B). Results are shown for both rVSV/SARS-CoV-2/GFP variants (the frequency of 1D7 mutations is plotted as circles and 2E1 mutations as triangles).

Figure 2—figure supplement 2
Analysis of S-encoding sequences following rVSV/SARS-CoV-2/GFP replication in the presence of convalescent plasma COV-107 and COV-NY.

(A–B) Graphs depict the S codon position (X-axis) and the frequency of non-synonymous substitutions (Y-axis) following the second, third or fourth passage (p2–p4) of rVSV/SARS-CoV-2/GFP on 293T/ACE2(B) cells in the presence of COV-107 plasma (A), or second passage in the presence of COV-NY plasma (B). Results are shown for both rVSV/SARS-CoV-2/GFP variants (the frequency of 1D7 mutations is plotted as circles and 2E1 mutations as triangles).

Figure 3 with 1 supplement
Characterization of mutant rVSV/SARS-CoV-2/GFP derivatives.

(A) Replication of plaque-purified rVSV/SARS-CoV-2/GFP bearing individual S amino-acid substitutions that arose during passage with the indicated antibody or plasma. 293T/ACE2cl.22 cells were inoculated with equivalent doses of parental or mutant rVSV/SARS-CoV-2/GFP isolates. Supernatant was collected at the indicated times after inoculation and number of infectious units present therein was determined on 293T/ACE2cl.22 cells. The mean of two independent experiments is plotted. One set of WT controls run concurrently with the mutants are replotted in the upper and lower left panels, A different set of WT controls run concurrently with the mutants is shown in the lower right panel (B) Infection 293T/ACE2cl.22 cells by rVSV/SARS-CoV-2/GFP encoding the indicated S protein mutations in the presence of increasing amounts of a chimeric ACE2-Fc molecule. Infection was quantified by FACS. Mean of two independent experiments is plotted. The WT controls are replotted in each panel.

Figure 3—figure supplement 1
Example of plaque purification of individual viral mutants from populations passaged in the presence of antibodies.

The upper panels show sequence traces from amplicons obtained from viral populations following replication in the presence of monoclonal antibodies, the bottom panels show sequence traces of amplicons obtained from mutants isolated by limiting dilution of the viral populations.

Neutralization of rVSV/SARS-CoV-2/GFP RBD mutants by monoclonal antibodies.

(A) Examples of neutralization resistance of rVSV/SARS-CoV-2/GFP mutants that were isolated following passage in the presence of antibodies. 293T/ACE2cl.22 cells were inoculated with WT or mutant rVSV/SARS-CoV-2/GFP in the presence of increasing amount of each monoclonal antibody, and infection quantified by FACS 16 hr later. Mean and SD from two technical replicates, representative of two independent experiments. (B) Neutralization sensitivity/resistance of rVSV/SARS-CoV-2/GFP mutants isolated following replication in the presence of antibodies. Mean IC50 values were calculated for each virus-monoclonal antibody combination in two independent experiments. (C) Position of neutralization resistance-conferring substitutions. Structure of the RBD (from PDB 6M17 Yan et al., 2020) with positions that are occupied by amino acids where mutations were acquired during replication in the presence of each monoclonal antibody or COV-NY plasma indicated.

Loss of binding to monoclonal antibodies by neutralization escape mutants.

(A) Schematic representation of the binding assay in which NanoLuc luciferase is appended to the C-termini of a conformationally stabilized S-trimer. The fusion protein is incubated with antibodies and complexes captured using protein G magnetic beads (B) Bound Nanoluc luciferase quantified following incubation of the indicated WT or mutant Nanoluc-S fusion proteins with the indicated antibodies and Protein G magnetic beads. Mean of three technical replicates at each S-Nanoluc concentration.

Neutralization of rVSV/SARS-CoV-2/GFP RBD mutants by convalescent plasma.

(A, B) Neutralization of rVSV/SARS-CoV-2/GFP mutants isolated following replication in the presence COV-47 plasma (A) or COV-NY plasma (B). 293T/ACE2cl.22 cells were inoculated with WT or mutant rVSV/SARS-CoV-2/GFP in the presence of increasing amounts of the indicated plasma, and infection quantified by flow cytometry, 16 hr later. Mean of two technical replicates, representative of two independent experiments (C) Plasma neutralization sensitivity/resistance of rVSV/SARS-CoV-2/GFP mutants isolated following replication in the presence of monoclonal antibodies or convalescent plasma. Mean NT50 values were calculated for each virus-plasma combination from two independent experiments.

Effects of naturally occurring RBD amino-acid substitutions on S sensitivity to neutralizing monoclonal antibodies.

(A–C) Neutralization of HIV-based reporter viruses pseudotyped with SARS-CoV-2 S proteins harboring the indicated naturally occurring substitutions. 293T/ACE2cl.22 cells were inoculated with equivalent doses of each pseudotyped virus in the presence of increasing amount of C121 (A) C135 (B) or C144 (C). Mean IC50 values were calculated for each virus-antibody combination from two independent experiments. (D) Position of substitutions conferring neutralization resistance relative to the amino acids close to the ACE2 binding site whose identity varies in global SARS-CoV-2 sequences. The RBD structure (from PDB 6M17 Yan et al., 2020) is depicted with naturally varying amino acids close to the ACE2 binding site colored in yellow. Amino acids whose substitution confers partial or complete (IC50 > 10 μg/ml) resistance to each monoclonal antibody in the HIV-pseudotype assays are indicated for C121 (red) C135 (green) and C144 (purple). (E) Binding of S-NanoLuc fusion protein in relative light units (RLU) to 293T or 293T/ACE2cl.22 cells after preincubation in the absence or presence of C121, C135, and C144 monoclonal antibodies. Each symbol represents a technical replicate.

Position and frequency of S amino-acid substitutions in SARS-CoV-2 S.

Global variant frequency reported by CoV-Glue in the SARS-CoV-2 S protein. Each individual variant is indicated by a symbol whose position in the S sequence is indicated on the X-axis and frequency reported by COV-Glue is indicated on the Y-axis. Individual substitutions at positions where mutations conferring resistance to neutralizing antibodies or plasma were found herein are indicated by enlarged and colored symbols: red for C121 and C144, green for C135, purple for COV-47 plasma and orange for COV-NY plasma. The common D/G614 variant is indicated.

Suppression of antibody resistance through the use of antibody combinations.

(A) Representative images of 293T/ACE2 (B) cells infected with the equivalent doses of rVSV/SARS-CoV-2/GFP in the absence or presence of 10 μg/ml of one (C144) or 5 μg/ml of each of two (C144 +C135) neutralizing monoclonal antibodies. (B) Expanded view of the boxed areas containing individual plaques from the culture infected in the presence of 10 μg/ml C144. (C) Expanded view of the boxed areas in A containing infected cells from the culture infected in the presence of 5 μg/ml each of (C144 and C135). (D) Infectious virus yield following two passages of rVSV/SARS-CoV-2/GFP in the absence or presence of individual neutralizing antibodies or combinations of two antibodies. Titers were determined on 293T/ACE2cl.22 cells. Each symbol represents a technical replicate and results from two independent experiments using rVSV/SARS-CoV-2/GFP1D7 and rVSV/SARS-CoV-2/GFP2E1 are shown.

Tables

Table 1
Plasma and monoclonal antibodies used in this study.
DonorPlasma NT50
(rVSV-SARSCoV2/GFP)
Plasma NT50
HIV/CCNGnLuc
Monoclonal antibody
COV-4766228016C144
COV-7262747982C135
COV-107122334C121
COV-NY126147505ND
Table 2
Mutations occurring at high frequency during rVSV/SARS-CoV-2 passage in the presence of neutralizing antibodies or plasma.
Mutant frequency
Mutationp2p3p4
Monoclonal antibodies
C121E484K*0.50, 0.39
F490L0.23
Q493K0.12, 0.45
C135N440K0.31, 0.30
R346S0.30, 0.17
R346K0.22, 0.53
R346M0.16
C144E484K0.44, 0.18
Q493K0.31, 0.39
Q493R0.17, 0.37
Plasmas
COV47N148S0.16, 0.140.29, 0.300.72, 0.14
K150R0.100.18
K150E0.040.160.4
K150T0.22
K150Q0.160.22
S151P0.10.180.2
COV-NYK444R0.20,0.19
K444N0.14
K444Q0.33
V445E0.18
  1. *Values represent the decimal frequency with which each mutation occurs ass assessed by NGS, two values indicate occurrences in both rVSV/SARS-CoV-2/GFP1D7 and rVSV/SARS-CoV-2/GFP2E1 cultures, single values indicate occurrence in only one culture.

    –, not done.

Key resources table
Reagent type
(species) or
resource
DesignationSource or
reference
IdentifiersAdditional
information
 Strain, strain background (Vesicular Stomatitis Virus)VSV/SARS-CoV-2/GFP1D7; WT1D7Schmidt et al., 2020Recombinant chimeric VSV/SARS-CoV-2 reporter virus
 Strain, strain background (Vesicular Stomatitis Virus)rVSV/SARS-CoV-2/GFP2E1; WT2E1Schmidt et al., 2020Recombinant chimeric VSV/SARS-CoV-2 reporter virus
 Strain, strain background (Vesicular Stomatitis Virus)E484K2E1Schmidt et al., 2020, and this papermutant rVSV/SARS-CoV-2/GFP derivative
Inquiries should be addressed to P.Bieniasz
 Strain, strain background (Vesicular Stomatitis Virus)Q493R1D7Schmidt et al., 2020, and this papermutant rVSV/SARS-CoV-2/GFP derivative
Inquiries should be addressed to P.Bieniasz
 Strain, strain background (Vesicular Stomatitis Virus)R346S1D7Schmidt et al., 2020, and this papermutant rVSV/SARS-CoV-2/GFP derivative
Inquiries should be addressed to P.Bieniasz
 Strain, strain background (Vesicular Stomatitis Virus)R3462E1Schmidt et al., 2020, and this papermutant rVSV/SARS-CoV-2/GFP derivative
Inquiries should be addressed to P.Bieniasz
 Strain, strain background (Vesicular Stomatitis Virus)N440K2E1Schmidt et al., 2020, and this papermutant rVSV/SARS-CoV-2/GFP derivative
Inquiries should be addressed to P.Bieniasz
 Strain, strain background (Vesicular Stomatitis Virus)K444N1D7Schmidt et al., 2020, and this papermutant rVSV/SARS-CoV-2/GFP derivative
Inquiries should be addressed to P.Bieniasz
 Strain, strain background (Vesicular Stomatitis Virus)K444T2E1Schmidt et al., 2020, and this papermutant rVSV/SARS-CoV-2/GFP derivative
Inquiries should be
addressed to P.Bieniasz
 Strain, strain background (Vesicular Stomatitis Virus)V445G2E1Schmidt et al., 2020, and this papermutant rVSV/SARS-CoV-2/GFP derivative
Inquiries should be addressed to P.Bieniasz
 Strain, strain background (Vesicular Stomatitis Virus)V445E1D7Schmidt et al., 2020, and this papermutant rVSV/SARS-CoV-2/GFP derivative
Inquiries should be addressed to P.Bieniasz
 Strain, strain background (Vesicular Stomatitis Virus)V445L2E1Schmidt et al., 2020, and this papermutant rVSV/SARS-CoV-2/GFP derivative
Inquiries should be addressed to P.Bieniasz
 Strain, strain background (Vesicular Stomatitis Virus)N148SSchmidt et al., 2020, and this papermutant rVSV/SARS-CoV-2/GFP derivative
Inquiries should be
addressed to P.Bieniasz
 Strain, strain background (Vesicular Stomatitis Virus)K150RSchmidt et al., 2020, and this papermutant rVSV/SARS-CoV-2/GFP derivative
Inquiries should be addressed to P.Bieniasz
 Strain, strain background (Vesicular Stomatitis Virus)K150ESchmidt et al., 2020, and this papermutant rVSV/SARS-CoV-2/GFP derivative
Inquiries should be addressed to P.Bieniasz
 Strain, strain background (Vesicular Stomatitis Virus)S151PSchmidt et al., 2020, and this papermutant rVSV/SARS-CoV-2/GFP derivative
Inquiries should be addressed to P.Bieniasz
 Cell line (Homo sapiens)Expi293F CellsThermo Fisher ScientificCat# A14527
 Cell line (H. sapiens)293T (embryonic, kidney)ATCCCRL-3216
 Cell line (H. sapiens)293T/ACE2(B)Schmidt et al., 2020293 T cells expressing human ACE2 (bulk population)
 Cell line (H. sapiens)293T/ACE2cl.22Schmidt et al., 2020293 T cells expressing human ACE2 (single cell clone)
 Biological sample (H. sapiens)COV-47Robbiani et al., 2020Human plasma sample
 Biological sample (H. sapiens)COV-72Robbiani et al., 2020Human plasma sample
 Biological sample (H. sapiens)COV-107Robbiani et al., 2020Human plasma sample
 Biological sample (H. sapiens)COV-NYLuchsinger et al., 2020Human plasma sample
 AntibodyC121
(Human monoclonal)
Robbiani et al., 2020Selection experiments (10 μg/ml, 5 μg/ml)
 AntibodyC135
(Human monoclonal)
Robbiani et al., 2020Selection experiments (10 μg/ml, 5 μg/ml)
 AntibodyC144
(Human monoclonal)
Robbiani et al., 2020Selection experiments (10 μg/ml, 5 μg/ml)
 Recombinant DNA reagentCSIB(ACE2)Schmidt et al., 2020
 Recombinant DNA reagentpHIVNLGagPolSchmidt et al., 2020
 Recombinant DNA reagentpCCNanoLuc2AEGFPSchmidt et al., 2020
 Recombinant DNA reagentpSARS-CoV-2Δ19Schmidt et al., 2020Epression plasmid containing a C-terminally truncated SARS-CoV-2 S protein (pSARS-CoV-2Δ19) containing a synthetic human-codon-optimized cDNA (Geneart)
 Recombinant DNA reagentR346SSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentR346KSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentV367FSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentN439KSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
rRcombinant DNA reagentN440KSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentK444QSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentK444RSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentK444NSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentV445ISchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentV445ESchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentV445LSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentV445KSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentG446VSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentG446SSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentL455RSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentL455ISchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentL455FSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentF456VSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentA475VSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentA475DSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentG476ASchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentG476SSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentT487ISchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentV483ISchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentV483ASchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentV483FSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentE484QSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentE484ASchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentE484DSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentF490SSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be
addressed to P.Bieniasz
Recombinant DNA reagentF490LSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant
DNA reagent
Q493KSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentQ493RSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentS494PSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentN501YSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Recombinant DNA reagentV503FSchmidt et al., 2020, and this paperpSARS-CoV-2Δ19
containing the indicated mutation.
Inquiries should be addressed to P.Bieniasz
Sequence-based reagentendof_M_forThis paperPCR and sequencing primerCTATCGGCCACTTCAAATGAGCTAG
Sequence-based reagentL_begin_revThis paperPCR and sequencing primerTCATGGAAGTCCACGATTTTGAGAC
Sequence-based reagentVSV-RBD-F primerThis paperPCR and sequencing primerCTGGCTCTGCACAGGTCCTACCTGACA
Sequence-based reagentVSV-RBD-R primerThis paperPCR and sequencing primerCAGAGACATTGTGTAGGCAATGATG
Peptide, recombinant proteinACE2-Fc fusion proteinThis paperRecombinant ACE2 extracellular domain fused to IgG1 Fc see Materials and Methods
Inquiries should be addressed to P.Bieniasz
Peptide, recombinant proteinS-6P-NanoLucThis paperA conformationally stabilized (6P) version of the SARS-CoV-2 S protein fused to Nanoluciferase See materials and methods
Inquiries should be addressed to P.Bieniasz
Commercial assay or kitTrizol-LSThermo FisherCat# 10296028
Commercial assay or kitSuperscript III reverse transcriptaseThermo FisherCat# 18080093
Commercial assay or kitNextera TDE1 Tagment DNA enzymeIlluminaCat# 150278650.25 µl
Commercial assay or kitTD Tagment DNA bufferIlluminaCat# 150278661.25 µl
commercial assay or kitNextera XT Index Kit v2IlluminaCat# FC-131–2001
Commercial assay or kitKAPA HiFi HotStart ReadyMixKAPA BiosystemsCat# KK2601
Commercial assay or kitAmPure Beads XPAgencourtCat# A63881
Commercial assay or kitExpi293 Expression System KitThermo Fisher ScientificCat# A14635
Commercial assay or kitNi-NTA AgaroseQiagenCat# 30210
Commercial assay or kitHRV 3C ProteaseTaKaRaCat# 7360
Commercial assay or kitLI-COR Intercept blocking bufferLicorP/N 927–70001
Commercial assay or kitDynabeads Protein GThermo Fisher ScientificCat# 10004D
Commercial assay or kitPassive Lysis 5X BufferPromegaCat# E1941
Commercial assay or kitNano-Glo Luciferase Assay SystemPromegaCat# N1150
Software, algorithmGeneious Primehttps://www.geneious.com/RRID:SCR_010519Version 2020.1.2
Software, algorithmPython programming languagehttps://www.python.org/
RRID:SCR_008394version 3.7
Software,
algorithm
pandas10.5281/zenodo.3509134RRID:SCR_018214Version 1.0.5
Software, algorithmnumpy10.1038/s41586-020-2649-2RRID:SCR_008633Version 1.18.5
Software, algorithmmatplotlib10.1109/MCSE.2007.55RRID:SCR_008624Version 3.2.2

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