An efficient approach for SARS-CoV-2 monoclonal antibody production via modified mRNA-LNP immunization

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1. Version published to 10.1016/j.ijpharm.2022.122256

   Nov 1, 2022
2. 

   ScreenIT

   Apr 25, 2022

   SciScore for 10.1101/2022.04.20.488878: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: Mouse immunization: All procedures involving animals were performed in accordance with guidelines set by the Institutional Animal Care and Use Committee (IACUC) at Academia Sinica, Taiwan. BALB/c mice 6 to 8 weeks of age were immunized by four intramuscular injections of 10 μg RBD mRNA-LNP at 2-week intervals or four intraperitoneal injections of 50 μg recombinant RBD protein in CFA-IFA adjuvant at 3-week intervals.
   Sex as a biological variable	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The plates were washed with PBST0.1 three times, then incubated with …

   SciScore for 10.1101/2022.04.20.488878: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: Mouse immunization: All procedures involving animals were performed in accordance with guidelines set by the Institutional Animal Care and Use Committee (IACUC) at Academia Sinica, Taiwan. BALB/c mice 6 to 8 weeks of age were immunized by four intramuscular injections of 10 μg RBD mRNA-LNP at 2-week intervals or four intraperitoneal injections of 50 μg recombinant RBD protein in CFA-IFA adjuvant at 3-week intervals.
   Sex as a biological variable	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The plates were washed with PBST0.1 three times, then incubated with horseradish peroxidase (HRP)-conjugated anti-human IgG (Sigma, MA, USA) secondary antibody at room temperature for 1 h.	anti-human IgG suggested: None
   Next, the horseradish peroxidase (HRP)-conjugated anti-mouse antibody (1:2000) was added for 1 h at room temperature.	anti-mouse suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   In vitro RBD expression of RBD mRNA and mRNA-LNPs: RBD mRNA was transfected into 293T cells via Lipofectamine™ 2000 Transfection Reagent (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA), while RBD mRNA-LNP was added directly to cells cultured in DMEM containing 10% FBS.	293T suggested: None
   Pseudovirus neutralization assay: SARS-CoV-2 pseudotyped lentiviruses expressing full-length S proteins and HEK293T cells overexpressing human ACE2 (HEK293T/hACE2) were provided by the National RNAi Core Facility (Academia Sinica, Taiwan).	HEK293T suggested: None
   The mixtures were added to pre-seeded Vero E6 cells for 1 h at 37°C.	Vero E6 suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Mouse immunization: All procedures involving animals were performed in accordance with guidelines set by the Institutional Animal Care and Use Committee (IACUC) at Academia Sinica, Taiwan. BALB/c mice 6 to 8 weeks of age were immunized by four intramuscular injections of 10 μg RBD mRNA-LNP at 2-week intervals or four intraperitoneal injections of 50 μg recombinant RBD protein in CFA-IFA adjuvant at 3-week intervals.	BALB/c suggested: None
   Software and Algorithms
   Sentences	Resources
   The hydrodynamic diameter (z-average) and zeta potential of RBD mRNA-LNPs were analyzed by Zetasizer software, version 7.11 (www.malvern.com).	Zetasizer suggested: None
   The half maximal inhibitory concentration (IC50) was calculated by nonlinear regression using Prism software version 8.1.0 (	Prism suggested: (PRISM, RRID:SCR_005375)
   GraphPad Software Inc.)	GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)

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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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   Results from scite Reference Check: We found no unreliable references.

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3. 

   Version published to 10.1101/2022.04.20.488878v1 on bioRxiv

   Apr 22, 2022