Yeast-expressed recombinant SARS-CoV-2 receptor binding domain RBD203-N1 as a COVID-19 protein vaccine candidate

Version published to 10.1016/j.pep.2021.106003

Feb 1, 2022

SciScore for 10.1101/2021.08.24.457518: (What is this?)

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

Table 1: Rigor

Ethics	IACUC: The reaction was terminated with 100 μL of 1M HCl and absorption readings were taken at 450 nm using a BioTek EPOCH 2 microplate reader. 2.10. Preclinical Study Design: A preclinical study in mice was performed under the approved Institutional Animal Care and Use Committee (IACUC) protocol at Baylor College of Medicine.
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
A 1:3,000 dilution of an AP-conjugated goat anti-mouse IgG (KPL) was used as the secondary antibody. 2.4. ELISA using Anti-RBD2l9-N1C1 Mouse Monoclonal Antibodies: In this …

SciScore for 10.1101/2021.08.24.457518: (What is this?)

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

Table 1: Rigor

Ethics	IACUC: The reaction was terminated with 100 μL of 1M HCl and absorption readings were taken at 450 nm using a BioTek EPOCH 2 microplate reader. 2.10. Preclinical Study Design: A preclinical study in mice was performed under the approved Institutional Animal Care and Use Committee (IACUC) protocol at Baylor College of Medicine.
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
A 1:3,000 dilution of an AP-conjugated goat anti-mouse IgG (KPL) was used as the secondary antibody. 2.4. ELISA using Anti-RBD2l9-N1C1 Mouse Monoclonal Antibodies: In this experiment, we evaluated the binding of eight anti-SARS-CoV-2 RBD219-WT mAbs (# 1128, 643, 486, 902, 854, 942, 748, and 102) to RBD203-N1 and RBD219-N1C1.	Anti-RBD2l9-N1C1 suggested: None anti-SARS-CoV-2 RBD219-WT suggested: None RBD219-N1C1 suggested: None
After this binding step, the plates were washed with PBST four times followed by adding 100 μL 1:6,000 diluted HRP conjugated anti-mouse IgG antibodies (LSBiosciences) and incubated for 1 hour at room temperature.	anti-mouse IgG suggested: None
After this binding step, the plates were washed with PBST four times followed by adding 100 μL of 1:5,000 diluted anti-RBD219-WT horse sera followed by 1:10,000 diluted HRP conjugated anti-horse IgG antibodies and incubating for 1 hour at room temperature.	anti-RBD219-WT suggested: None anti-horse IgG suggested: None
6–8-week-old Female BALB/c mice were immunized twice intramuscularly (i.m.) at 21-day intervals and then euthanized 14 days after the second immunization. 2.11. Antigen-specific Antibody Measurements by ELISA: To examine RBD-specific antibodies in mouse sera, indirect ELISAs were conducted as described previously [13].	Antigen-specific suggested: None
Experimental Models: Cell Lines
Sentences	Resources
Next, 100 μL of sera-pseudovirus were added to 293T-hACE2 cells in 96-well poly-D-lysine coated culture plates.	293T-hACE2 suggested: None
Experimental Models: Organisms/Strains
Sentences	Resources
6–8-week-old Female BALB/c mice were immunized twice intramuscularly (i.m.) at 21-day intervals and then euthanized 14 days after the second immunization. 2.11. Antigen-specific Antibody Measurements by ELISA: To examine RBD-specific antibodies in mouse sera, indirect ELISAs were conducted as described previously [13].	BALB/c suggested: None
Recombinant DNA
Sentences	Resources
In short, the DNA encoding RBD203-N1 was synthesized and subcloned into the Pichia secretory expression vector pPICZαA (Invitrogen) using EcoRI/Xbal restriction sites (GenScript).	pPICZαA suggested: None
The RBD203-N1 pPICZαA/P. pastoris X33 construct was fermented in 5 L vessels [9, 11, 12].	pPICZαA/P suggested: None
The plasmids used for the pseudovirus production are the luciferase-encoding reporter plasmid (pNL4-3.lucR-E-),	pNL4-3.lucR-E- suggested: None
, Gag/Pol-encoding packaging construct (pΔ8.9), and codon-optimized SARS-CoV-2 spike protein expression plasmids (pcDNA3.1-CoV-2 S gene) based on clone p278-1.	pcDNA3.1-CoV-2 suggested: None
Software and Algorithms
Sentences	Resources
Raw data were first analyzed by Bio-Plex Manager software followed by Excel and Prism. 2.13.	Bio-Plex Manager suggested: None Excel suggested: None Prism suggested: (PRISM, RRID:SCR_005375)

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).

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.

Results from TrialIdentifier: No clinical trial numbers were referenced.

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

Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on page 23. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

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.

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

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Yeast-expressed recombinant SARS-CoV-2 receptor binding domain RBD203-N1 as a COVID-19 protein vaccine candidate

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