Optimization of an LNP-mRNA vaccine candidate targeting SARS-CoV-2 receptor-binding domain

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   ScreenIT

   Mar 15, 2021

   SciScore for 10.1101/2021.03.04.433852: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: All mouse studies were approved by the Animal Experiment Committee of the Institute of Medical Science, University of Tokyo. Consent: Peripheral blood mononuclear cells (PBMCs) were obtained from three SARS-CoV-2-uninfected healthy adult volunteers after obtaining informed consent. IRB: All experiments using human PBMCs were approved by the Institutional Review Board of the Institute of Medical Science, University of Tokyo.
   Randomization	not detected.
   Blinding	Histological evaluation was performed blindly by two pathologists based on a following criteria established in influenza virus infection 26 (0: normal lung, 1: mild destruction of bronchial epithelium, 2: …

   SciScore for 10.1101/2021.03.04.433852: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: All mouse studies were approved by the Animal Experiment Committee of the Institute of Medical Science, University of Tokyo. Consent: Peripheral blood mononuclear cells (PBMCs) were obtained from three SARS-CoV-2-uninfected healthy adult volunteers after obtaining informed consent. IRB: All experiments using human PBMCs were approved by the Institutional Review Board of the Institute of Medical Science, University of Tokyo.
   Randomization	not detected.
   Blinding	Histological evaluation was performed blindly by two pathologists based on a following criteria established in influenza virus infection 26 (0: normal lung, 1: mild destruction of bronchial epithelium, 2: mild peribronchiolar inflammation, 3: inflammation in the alveolar walls resulting in alveolar thickening, 4: mild alveolar injury accompanied by vascular injury, 5: moderate alveolar injury and vascular injury, 6, 7: severe alveolar injury with hyaline membrane-associated alveolar hemorrhage (under or over 50% of the section area)).
   Power Analysis	not detected.
   Sex as a biological variable	Cynomolgus macaque: Seven to ten-year-old female cynomolgus macaques born at Shiga University of Medical Science and originating from Philippines, Vietnam, and China were used.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   , anti-CD4 (RM4-5), anti-CD185 (L138D7), anti-CD38 (90), and anti-CD19 (6D5) antibodies.	anti-CD4 suggested: (BD Biosciences Cat# 340573, RRID:AB_400476) anti-CD185 suggested: None anti-CD38 suggested: (BD Biosciences Cat# 341132, RRID:AB_647410) anti-CD19 ( 6D5 ) suggested: None
   , anti-CD19 (6D5), anti-CD11c (N418), anti-NK-1.1 (PK136), and anti-CD45R/B220 (RA3-6B2) antibodies.	anti-CD19 suggested: None anti-CD11c suggested: None anti-NK-1.1 suggested: None anti-CD45R/B220 suggested: (STEMCELL Technologies Cat# 10712, RRID:AB_215616) RA3-6B2 suggested: None
   , anti-TNF-α (MP6-XT22), and anti-CD3 (17A2) antibodies.	anti-TNF-α suggested: (Leinco Technologies Cat# T798, RRID:AB_2832121) anti-CD3 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Virus: SARS-CoV-2 isolates were propagated in VeroE6 cells in Opti-MEM I (Invitrogen, Carlsbad, CA, USA) containing 0.3% bovine serum albumin (BSA) and 1 μg of L-1-tosylamide-2-phenylethyl chloromethyl ketone (TPCK)-treated trypsin/mL at 37°C. Immunization: Six to eight week-old C57BL/6 and BALB/c mice were intramuscularly immunized with mock, LNP-mRNA-RBD (3 μg), or LNP-mRNA-RBD (HPLC) (3 μg) on days 0 and 14.	VeroE6 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
   Neutralization activity against SARS-CoV-2 infection: Thirty-five microliters of virus (140 tissue culture infectious dose 50) was incubated with 35 μL of two-fold serial dilutions of sera for 1 h at room temperature, and 50 μL of the mixture was added to confluent VeroE6/TMPRSS2 cells in 96-well plates and incubated for 1 h at 37°C.	VeroE6/TMPRSS2 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Mice: Six to eight week-old C57BL/6 and BALB/c mice were purchased from CLEA, Japan.	C57BL/6 suggested: None
   Virus: SARS-CoV-2 isolates were propagated in VeroE6 cells in Opti-MEM I (Invitrogen, Carlsbad, CA, USA) containing 0.3% bovine serum albumin (BSA) and 1 μg of L-1-tosylamide-2-phenylethyl chloromethyl ketone (TPCK)-treated trypsin/mL at 37°C. Immunization: Six to eight week-old C57BL/6 and BALB/c mice were intramuscularly immunized with mock, LNP-mRNA-RBD (3 μg), or LNP-mRNA-RBD (HPLC) (3 μg) on days 0 and 14.	BALB/c suggested: None

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   Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

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   Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

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   Results from JetFighter: We did not find any issues relating to colormaps.

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   Results from rtransparent:

   • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
   • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
   • No protocol registration statement was detected.

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   Version published to 10.1101/2021.03.04.433852v1 on bioRxiv

   Mar 4, 2021