Next generation vaccine platform: polymersomes as stable nanocarriers for a highly immunogenic and durable SARS-CoV-2 spike protein subunit vaccine

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   ScreenIT

   Jan 26, 2021

   SciScore for 10.1101/2021.01.24.427729: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement

   IACUC: All experiments and procedures were approved by the Institutional Animal Care and Use Committee of the Biological Resource Center (Agency for Science, Technology and Research, Singapore) in accordance with the guidelines of the Agri-Food and Veterinary Authority and the National Advisory Committee for Laboratory Animal Research of Singapore (ICUAC No. 181357). Consent: ACM uptake in human PBMC: Blood samples were obtained from healthy donors after provided written informed consent to participate in research protocols approved by the Institutional Review Board of Singapore Immunology Network (SIgN) IRB: ACM uptake in human PBMC: Blood samples were obtained …

   SciScore for 10.1101/2021.01.24.427729: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: All experiments and procedures were approved by the Institutional Animal Care and Use Committee of the Biological Resource Center (Agency for Science, Technology and Research, Singapore) in accordance with the guidelines of the Agri-Food and Veterinary Authority and the National Advisory Committee for Laboratory Animal Research of Singapore (ICUAC No. 181357). Consent: ACM uptake in human PBMC: Blood samples were obtained from healthy donors after provided written informed consent to participate in research protocols approved by the Institutional Review Board of Singapore Immunology Network (SIgN) IRB: ACM uptake in human PBMC: Blood samples were obtained from healthy donors after provided written informed consent to participate in research protocols approved by the Institutional Review Board of Singapore Immunology Network (SIgN)
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Female C57BL/6 mice were purchased from InVivos and used at 8-9 weeks of age.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The membrane was washed thrice with TBST for a total of 30 min before incubating 1 h at room temperature with HRP-conjugated goat anti-mouse secondary antibody at a 1:10,000 dilution.	anti-mouse suggested: None
   Monoclonal antibodies against Ly6C (clone HK1.4), CD11b (clone M1/70),	Ly6C suggested: None CD11b suggested: None
   CD49b (clone HMa2), and Ly6G (clone 1A8) were purchased from BD Bioscience, CD19 (clone 1D3) and Streptavidin for conjugation of biotinylated antibodies were purchased from BD Horizon.	Ly6G suggested: None CD19 suggested: None
   Splenocytes were incubated with an overlapping peptide pool covering the spike protein (JPT product PM-WCPV-S-1 Vials 1 and 2) along with functional anti-mouse CD28 and CD49d antibodies overnight at 37°C, 5% CO2.	anti-mouse CD28 suggested: None CD49d suggested: None
   PBMCs were stained with mouse anti-human monoclonal surface antibodies (mAbs) against CD45 (clone HI30),	anti-human monoclonal surface antibodies suggested: None CD45 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   The gene of interest was placed into the Bac-to-Bac system (Thermo Fisher Scientific), transfected and passaged in Sf9 cells (Thermo Fisher Scientific) until a high titre was achieved.	Sf9 suggested: CLS Cat# 604328/p700_Sf9, RRID:CVCL_0549)
   Pseudovirus neutralization test: Pseudotyped lentiviral particles harbouring the SARS-CoV-2 spike glycoprotein (S-pp) were generated by co-transfection of 293FT cells with S expression plasmid and envelope-defective pNL4-3.	293FT suggested: ATCC Cat# PTA-5077, RRID:CVCL_6911)
   CHO cells stably overexpressing human ACE2 (CHO-ACE2) (55) were seeded in 96-well plates 24 hour before transduction.	CHO suggested: None
   The mixture was transferred to Vero-E6 cells and incubated for 1 h at 37°C.	Vero-E6 suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Mice (investigation of DC targeting by ACM polymersomes): C57BL/6 mice were purchased from InVivos.	C57BL/6 suggested: None
   Software and Algorithms
   Sentences	Resources
   The trimeric spike protein was purchased from ACROBiosystems and the S2 domain protein from Sino Biological.	ACROBiosystems suggested: (ACRObiosystems, RRID:SCR_012550)
   Images were analyzed in Fiji ImageJ software (v.	ImageJ suggested: (ImageJ, RRID:SCR_003070)
   Flow cytometry acquisition was performed on a 5-laser LSR II (BD) using FACSDiva software, and data subsequently analyzed with FlowJo v.	FACSDiva suggested: (BD FACSDiva Software, RRID:SCR_001456)
   Data were analyzed using FlowJo v.	FlowJo suggested: (FlowJo, RRID:SCR_008520)
   Background absorbance was subtracted and the EC50 value of the titration curve was determined using GraphPad Prism version 8.4.3 with five-parameter non-linear regression.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)
   Each titration curve was analysed via five-parameter non-linear regression (GraphPad Prism V8.4.3) to calculate endpoint titer, which was defined as the highest dilution producing an absorbance three times the plate background.	GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)
   The S expression plasmid was constructed by cloning the codon-optimised spike gene (according to GenBank accession QHD43416.1) containing a 19 amino acid C-terminal truncation to enhance pseudotyping efficiency (36) into the pTT5 mammalian expression vector (pTT5LnX-coV-SP, a kind gift from Brendon John Hanson, Biological Defence Program, DSO National Laboratories, Singapore).	Biological Defence Program suggested: (UniProtKB, RRID:SCR_004426)

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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: Please consider improving the rainbow (“jet”) colormap(s) used on page 30. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

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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 10.1101/2021.01.24.427729v1 on bioRxiv

   Jan 25, 2021