COVID-19 in Tunisia (North Africa): IgG and IgG subclass antibody responses to SARS-CoV-2 according to disease severity

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1. 

   ScreenIT

   Mar 5, 2022

   SciScore for 10.1101/2022.03.01.22271696: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: The study protocol was approved by the Institut Pasteur de Tunis ethical committee (2020/21/I/LR16IPT/V2). Consent: Patients were included after written informed consent.
   Sex as a biological variable	Non-inclusion criteria were as follows: presence of a mental handicap, pregnant women and patients on immunosuppressive therapy.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The quality control of the produced recombinant proteins was performed using several tests including western blot and MALDI-TOF. 2.3 Indirect ELISA for IgG antibodies to S-RBD and N SARS COV 2 proteins: …

   SciScore for 10.1101/2022.03.01.22271696: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: The study protocol was approved by the Institut Pasteur de Tunis ethical committee (2020/21/I/LR16IPT/V2). Consent: Patients were included after written informed consent.
   Sex as a biological variable	Non-inclusion criteria were as follows: presence of a mental handicap, pregnant women and patients on immunosuppressive therapy.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The quality control of the produced recombinant proteins was performed using several tests including western blot and MALDI-TOF. 2.3 Indirect ELISA for IgG antibodies to S-RBD and N SARS COV 2 proteins: Ninety-six–well ELISA plates were coated overnight with recombinant proteins in phosphate-buffered saline (PBS) (50 ng for N and 100 ng for S-RBD in a volume of 50 µl per well).	IgG suggested: None
   2.4 Indirect ELISA for IgG subclass antibodies to S-RBD and N SARS COV 2 proteins: Plates were coated overnight with recombinant N protein (1µg/ml) or S-RBD (2µg/ml).	IgG subclass suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Sf-9 cells were then infected with the recombinant baculoviruses at an MOI of 3.	Sf-9 suggested: CLS Cat# 604328/p700_Sf9, RRID:CVCL_0549)
   Plates were then washed six times, incubated with 8000-fold diluted peroxidase-conjugated goat anti-human IgG (Sigma, AG029) for 1 hour at 37°C.	AG029 suggested: None
   Recombinant DNA
   Sentences	Resources
   2.2 Recombinant proteins production and purification: Recombinant N protein was produced in Escherichia coli BL21 (DE3) using the pETM11/N-nCov-(His)6-Nter plasmid as previously described (Grzelak et al. 2020b).	pETM11/N-nCov- suggested: None
   The S-RBD (residues 319–541) of the SARS-CoV-2 spike protein (Wang et al. 2020) was cloned into a customized pFastBac vector.	pFastBac suggested: RRID:Addgene_1925)
   Software and Algorithms
   Sentences	Resources
   Statistical analyses using nonparametric tests were performed using GraphBad Prism version 5 and p<0.05 was considered to be statistically significant.	GraphBad Prism 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: We detected the following sentences addressing limitations in the study:

   Our study has some limitations. We did not evaluate the effects of gender and age on antibodies responses in association with COVID-19 severity nor did we test to what extent comorbidities may impact the immune response against SARS-CoV-2. Further longitudinal studies are needed to monitor how the immune responses of patients with COVID-19 evolve over time in order to assess the immune signatures that may herald either recovery or aggravation.

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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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2. 

   Version published to 10.1101/2022.03.01.22271696v1 on medRxiv

   Mar 2, 2022