Effective screening of SARS-CoV-2 neutralizing antibodies in patient serum using lentivirus particles pseudotyped with SARS-CoV-2 spike glycoprotein
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Abstract
Pseuodotyped particles have significant importance and use in virology as tools for studying the biology of highly pathogenic viruses in a lower biosafety environment. The biological, chemical, and serological studies of the recently emerged SARS-CoV-2 will be greatly aided by the development and optimization of a suitable pseudotyping system. Here, we pseudotyped the SARS-CoV-2 Spike glycoprotein (SPG) on a traditional retroviral (MMLV) as well as a third generation lentiviral (pLV) vector and tested the transduction efficiency in several mammalian cell lines expressing SARS-CoV-2 receptor hACE2. While MMLV pseudotyped the vesicular stomatitis virus G glycoprotein (VSV-G) efficiently, it could not pseudotype the full-length SPG. In contrast, pLV pseudotyped both glycoproteins efficiently; however, much higher titers of pLV-G particles were produced. Among all the tested mammalian cells, 293Ts expressing hACE2 were most efficiently transduced using the pLV-S system. The pLV-S particles were efficiently neutralized by diluted serum (>:640) from recently recovered COVID-19 patients who showed high SARS-CoV-2 specific IgM and IgG levels. In summary, pLV-S pseudotyped virus provides a valid screening tool for the presence of anti SARS-CoV-2 specific neutralizing antibodies in convalescent patient serum.
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SciScore for 10.1101/2020.05.21.20108951: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: Written informed consent was obtained from the individuals recruited for this study.
IRB: The study was reviewed and approved by the Institutional Review Board of the University of Mississippi Medical Center which follows the national and international guidelines consistent with the principles established by the Declaration of Helsinki.Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Cells: 293T (ATCC # CRL3216), Vero E6 (ATCC # CRL1586) and MDCK (ATCC #CCL-34) were cultured in DMEM … SciScore for 10.1101/2020.05.21.20108951: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: Written informed consent was obtained from the individuals recruited for this study.
IRB: The study was reviewed and approved by the Institutional Review Board of the University of Mississippi Medical Center which follows the national and international guidelines consistent with the principles established by the Declaration of Helsinki.Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Cells: 293T (ATCC # CRL3216), Vero E6 (ATCC # CRL1586) and MDCK (ATCC #CCL-34) were cultured in DMEM (Corning Inc) supplemented with 10% fetal bovine serum (FBS, Fisher Scientific). 293Tsuggested: ATCC Cat# CRL-3216, RRID:CVCL_0063)Vero E6suggested: NoneMDCKsuggested: ATCC Cat# CCL-34, RRID:CVCL_0422)A549 (ATCC #CCL-185) were cultured in F-12K (ATCC #30-2004) supplemented with 10% fetal bovine serum. A549suggested: NoneHEK293T cells were transfected with the following plasmids:10 μg pLNCX-GFP, 1.2 μg pCMV-tat-HIV, 6 μg pJK3, and 3 μg of pL-VSV-G (control) or pCAGGS-S (SARS-CoV-2). HEK293Tsuggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)Recombinant DNA Sentences Resources 293T cells were transfected with hACE2 plasmid (a gift from Hyeryun Choe (Addgene plasmid # 1786; http://n2t.net/addgene:1786; RRID:Addgene_1786) to produce hACE2-293T cells. detected: RRID:Addgene_1786)Generation of pseudotype particles: HEK293T cells (2 x 106) were plated in a 100-mm tissue culture dish and transfected the next day when they were about 75% confluent with a combination of the following plasmids: 9 μg of pLV-eGFP (a gift from Pantelis Tsoulfas (Addgene plasmid # 36083; http://n2t.net/addgene:36083; RRID:Addgene_36083), 9 μg of psPAX2 (a gift from Didier Trono (Addgene plasmid # 12260; http://n2t.net/addgene:12260; RRID:Addgene_12260), and 3 μg of pCAGGS-S (SARS-CoV-2)(Catalog No. NR-52310:BEI Resources) or VSV-G (a gift from Tannishtha Reya (Addgene plasmid # 14888; http://n2t.net/addgene:14888; RRID:Addgene_14888) as control. detected: RRID:Addgene_36083)detected: RRID:Addgene_12260)detected: RRID:Addgene_14888)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: We did not find any issues relating to colormaps.
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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SciScore for 10.1101/2020.05.21.20108951: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Written informed consent was obtained from the individuals recruited for this study. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Methods Cells: 293T (ATCC # CRL3216), Vero E6 (ATCC # CRL1586) and MDCK (ATCC #CCL-34) were cultured in DMEM (Corning Inc) supplemented with 10% fetal bovine serum (FBS, Fisher Scientific). …293Tsuggested: ATCC Cat# CRL-3216, RRID:CVCL_0063SciScore for 10.1101/2020.05.21.20108951: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Written informed consent was obtained from the individuals recruited for this study. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Methods Cells: 293T (ATCC # CRL3216), Vero E6 (ATCC # CRL1586) and MDCK (ATCC #CCL-34) were cultured in DMEM (Corning Inc) supplemented with 10% fetal bovine serum (FBS, Fisher Scientific). 293Tsuggested: ATCC Cat# CRL-3216, RRID:CVCL_0063Vero E6suggested: NoneMDCKsuggested: ATCC Cat# CCL-34, RRID:CVCL_0422A549 (ATCC #CCL-185) were cultured in F-12K (ATCC #30-2004) supplemented with 10% fetal bovine serum. A549suggested: NoneGeneration of pseudotype particles: HEK293T cells (2 x 106) were plated in a 100-mm tissue culture dish and transfected the next day when they were about 75% confluent w combination of the following plasmids: 9 µg of pLV-eGFP (a gift from Pantelis Tsoulfas (Addgene plasmid # 36083 ; http://n2t.net/addgene:36083 ; RRID:Addgene_36083), 9 µ psPAX2 (a gift from Didier Trono (Addgene plasmid # 12260 ; http://n2t.net/addgene:12 RRID:Addgene_12260), and 3 µg of pCAGGS-S (SARS-CoV-2)(Catalog No. NR-52310 Resources) or VSV-G (a gift from Tannishtha Reya (Addgene plasmid # 14888 ; http://n2t.net/addgene:14888 ; RRID:Addgene_14888) as control. HEK293Tsuggested: None), adenocarcinomic human alveolar ar basal epithelial cells (A549), and Vero (VeroE6) cells depicted as a percentage of efficiency in 293T cells transfected with plasmid encoding hACE2 receptor (hACE2293T). Verosuggested: NoneRecombinant DNA Sentences Resources 293T cells were transfected with hACE2 plasmid (a gift from Hyeryun Choe (Addgene plasmid # 1786 ; http://n2t.net/addgene:1786 ; RRID:Addgene_1786) to produce hACE2-293T cells. detected: RRID:Addgene_1786Generation of pseudotype particles: HEK293T cells (2 x 106) were plated in a 100-mm tissue culture dish and transfected the next day when they were about 75% confluent w combination of the following plasmids: 9 µg of pLV-eGFP (a gift from Pantelis Tsoulfas (Addgene plasmid # 36083 ; http://n2t.net/addgene:36083 ; RRID:Addgene_36083), 9 µ psPAX2 (a gift from Didier Trono (Addgene plasmid # 12260 ; http://n2t.net/addgene:12 RRID:Addgene_12260), and 3 µg of pCAGGS-S (SARS-CoV-2)(Catalog No. NR-52310 Resources) or VSV-G (a gift from Tannishtha Reya (Addgene plasmid # 14888 ; http://n2t.net/addgene:14888 ; RRID:Addgene_14888) as control. detected: RRID:Addgene_36083detected: RRID:Addgene_12260detected: RRID:Addgene_14888Results 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 Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).
Results from JetFighter: We did not find any issues relating to colormaps.
About SciScore
SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore is not a substitute for expert review. SciScore checks for the presence and correctness of RRIDs (research resource identifiers) in the manuscript, and detects sentences that appear to be missing RRIDs. SciScore also checks to make sure that rigor criteria are addressed by authors. It does this by detecting sentences that discuss criteria such as blinding or power analysis. SciScore does not guarantee that the rigor criteria that it detects are appropriate for the particular study. Instead it assists authors, editors, and reviewers by drawing attention to sections of the manuscript that contain or should contain various rigor criteria and key resources. For details on the results shown here, including references cited, please follow this link.
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SciScore for 10.1101/2020.05.21.20108951: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Written informed consent was obtained from the individuals recruited for this study. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Methods Cells: 293T (ATCC # CRL3216), Vero E6 (ATCC # CRL1586) and MDCK (ATCC #CCL-34) were cultured in DMEM (Corning Inc) supplemented with 10% fetal bovine serum (FBS, Fisher Scientific). Vero E6detected: None…SciScore for 10.1101/2020.05.21.20108951: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Written informed consent was obtained from the individuals recruited for this study. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Methods Cells: 293T (ATCC # CRL3216), Vero E6 (ATCC # CRL1586) and MDCK (ATCC #CCL-34) were cultured in DMEM (Corning Inc) supplemented with 10% fetal bovine serum (FBS, Fisher Scientific). Vero E6detected: NoneMDCKsuggested: ATCC Cat# CCL-34, RRID:CVCL_0422A549 (ATCC #CCL-185) were cultured in F-12K (ATCC #30-2004) supplemented with 10% fetal bovine serum. A549detected: None293T cells were transfected with hACE2 plasmid (a gift from Hyeryun Choe (Addgene plasmid # 1786 ; http://n2t.net/addgene:1786 ; RRID:Addgene_1786) to produce hACE2-293T cells. 293Tdetected: None(E) Transduction efficiency of pLV pseudotyped with CoV-2 Spike glycoprotein in HEK293T, Madin-Darby Canine Kidney cells (MDCK) HEK293Tsuggested: KCB Cat# KCB 200744YJ, RRID:CVCL_0063), adenocarcinomic human alveolar ar basal epithelial cells (A549), and Vero (VeroE6) cells depicted as a percentage of efficiency in 293T cells transfected with plasmid encoding hACE2 receptor (hACE2293T). Verodetected: NoneRecombinant DNA Sentences Resources 293T cells were transfected with hACE2 plasmid (a gift from Hyeryun Choe (Addgene plasmid # 1786 ; http://n2t.net/addgene:1786 ; RRID:Addgene_1786) to produce hACE2-293T cells. detected: RRID:Addgene_1786Generation of pseudotype particles: HEK293T cells (2 x 106) were plated in a 100-mm tissue culture dish and transfected the next day when they were about 75% confluent w combination of the following plasmids: 9 µg of pLV-eGFP (a gift from Pantelis Tsoulfas (Addgene plasmid # 36083 ; http://n2t.net/addgene:36083 ; RRID:Addgene_36083), 9 µ psPAX2 (a gift from Didier Trono (Addgene plasmid # 12260 ; http://n2t.net/addgene:12 RRID:Addgene_12260), and 3 µg of pCAGGS-S (SARS-CoV-2)(Catalog No. NR-52310 Resources) or VSV-G (a gift from Tannishtha Reya (Addgene plasmid # 14888 ; http://n2t.net/addgene:14888 ; RRID:Addgene_14888) as control. detected: RRID:Addgene_36083detected: RRID:Addgene_12260detected: RRID:Addgene_14888Results 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 Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).
Results from JetFighter: We did not find any issues relating to colormaps.
About SciScore
SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore is not a substitute for expert review. SciScore checks for the presence and correctness of RRIDs (research resource identifiers) in the manuscript, and detects sentences that appear to be missing RRIDs. SciScore also checks to make sure that rigor criteria are addressed by authors. It does this by detecting sentences that discuss criteria such as blinding or power analysis. SciScore does not guarantee that the rigor criteria that it detects are appropriate for the particular study. Instead it assists authors, editors, and reviewers by drawing attention to sections of the manuscript that contain or should contain various rigor criteria and key resources. For details on the results shown here, including references cited, please follow this link.
-
SciScore for 10.1101/2020.05.21.20108951: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Written informed consent was obtained from the individuals recruited for this study. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Serial dilutions (1:40, 1:80, 1:160: 1:320, 1:640 and 1:1280) of serum were made in the serum-free cell culture medium and incubated with equal amounts of pLV-S virus, then plated on 293T cells. 293Tsuggested: KCB Cat# KCB 200744YJ, CVCL_0063Metho… SciScore for 10.1101/2020.05.21.20108951: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Written informed consent was obtained from the individuals recruited for this study. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Serial dilutions (1:40, 1:80, 1:160: 1:320, 1:640 and 1:1280) of serum were made in the serum-free cell culture medium and incubated with equal amounts of pLV-S virus, then plated on 293T cells. 293Tsuggested: KCB Cat# KCB 200744YJ, CVCL_0063Methods Cells: 293T (ATCC # CRL3216), Vero E6 (ATCC # CRL1586) and MDCK (ATCC #CCL-34) were cultured in DMEM (Corning Inc) supplemented with 10% fetal bovine serum (FBS, Fisher Scientific). Vero E6detected: NoneMDCKsuggested: ATCC Cat# CCL-34, CVCL_0422A549 (ATCC #CCL-185) were cultured in F-12K (ATCC #30-2004) supplemented with 10% fetal bovine serum. A549detected: None(E) Transduction efficiency of pLV pseudotyped with CoV-2 Spike glycoprotein in HEK293T, Madin-Darby Canine Kidney cells (MDCK) HEK293Tsuggested: KCB Cat# KCB 200744YJ, CVCL_0063), adenocarcinomic human alveolar ar basal epithelial cells (A549), and Vero (VeroE6) cells depicted as a percentage of efficiency in 293T cells transfected with plasmid encoding hACE2 receptor (hACE2293T). Verodetected: NoneRecombinant DNA Sentences Resources 293T cells were transfected with hACE2 plasmid (a gift from Hyeryun Choe (Addgene plasmid # 1786 ; http://n2t.net/addgene:1786 ; Addgene_1786) to produce hACE2-293T cells. detected: Addgene_1786Generation of pseudotype particles: HEK293T cells (2 x 106) were plated in a 100-mm tissue culture dish and transfected the next day when they were about 75% confluent w combination of the following plasmids: 9 µg of pLV-eGFP (a gift from Pantelis Tsoulfas (Addgene plasmid # 36083 ; http://n2t.net/addgene:36083 ; Addgene_36083), 9 µ psPAX2 (a gift from Didier Trono (Addgene plasmid # 12260 ; http://n2t.net/addgene:12 Addgene_12260), and 3 µg of pCAGGS-S (SARS-CoV-2)(Catalog No. NR-52310 Resources) or VSV-G (a gift from Tannishtha Reya (Addgene plasmid # 14888 ; http://n2t.net/addgene:14888 ; Addgene_14888) as control. detected: Addgene_36083detected: Addgene_12260detected: Addgene_14888Results 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 Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).
Results from JetFighter: We did not find any issues relating to colormaps.
About SciScore
SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore is not a substitute for expert review. SciScore checks for the presence and correctness of RRIDs (research resource identifiers) in the manuscript, and detects sentences that appear to be missing RRIDs. SciScore also checks to make sure that rigor criteria are addressed by authors. It does this by detecting sentences that discuss criteria such as blinding or power analysis. SciScore does not guarantee that the rigor criteria that it detects are appropriate for the particular study. Instead it assists authors, editors, and reviewers by drawing attention to sections of the manuscript that contain or should contain various rigor criteria and key resources. For details on the results shown here, including references cited, please follow this link.
-
SciScore for 10.1101/2020.05.21.20108951: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Written informed consent was obtained from the individuals recruited for this study. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Methods Cells: 293T (ATCC # CRL3216), Vero E6 (ATCC # CRL1586) and MDCK (ATCC #CCL-34) were cultured in DMEM (Corning Inc) supplemented with 10% fetal bovine serum (FBS, Fisher Scientific). …293Tsuggested: ATCC Cat# CRL-3216, CVCL_0063SciScore for 10.1101/2020.05.21.20108951: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Written informed consent was obtained from the individuals recruited for this study. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Experimental Models: Cell Lines Sentences Resources Methods Cells: 293T (ATCC # CRL3216), Vero E6 (ATCC # CRL1586) and MDCK (ATCC #CCL-34) were cultured in DMEM (Corning Inc) supplemented with 10% fetal bovine serum (FBS, Fisher Scientific). 293Tsuggested: ATCC Cat# CRL-3216, CVCL_0063Vero E6detected: NoneA549 (ATCC #CCL-185) were cultured in F-12K (ATCC #30-2004) supplemented with 10% fetal bovine serum. A549detected: NoneHEK293T were transfected with the following plasmids: 10 µg pLNCX-GFP, 1.2 µg pCMV-tat-HIV, pJK3, and 3 µg of pL-VSV-G (control) or pCAGGS-S (SARS-CoV-2). HEK293Tsuggested: KCB Cat# KCB 200744YJ, CVCL_0063(E) Transduction efficiency of pLV pseudotyped with CoV-2 Spike glycoprotein in HEK293T, Madin-Darby Canine Kidney cells (MDCK) MDCKsuggested: CLS Cat# 602280/p823_MDCK_(NBL-2), CVCL_0422), adenocarcinomic human alveolar ar basal epithelial cells (A549), and Vero (VeroE6) cells depicted as a percentage of efficiency in 293T cells transfected with plasmid encoding hACE2 receptor (hACE2293T). Verodetected: NoneRecombinant DNA Sentences Resources 293T cells were transfected with hACE2 plasmid (a gift from Hyeryun Choe (Addgene plasmid # 1786 ; http://n2t.net/addgene:1786 ; Addgene_1786) to produce hACE2-293T cells. detected: Addgene_1786Generation of pseudotype particles: HEK293T cells (2 x 106) were plated in a 100-mm tissue culture dish and transfected the next day when they were about 75% confluent w combination of the following plasmids: 9 µg of pLV-eGFP (a gift from Pantelis Tsoulfas (Addgene plasmid # 36083 ; http://n2t.net/addgene:36083 ; Addgene_36083), 9 µ psPAX2 (a gift from Didier Trono (Addgene plasmid # 12260 ; http://n2t.net/addgene:12 Addgene_12260), and 3 µg of pCAGGS-S (SARS-CoV-2)(Catalog No. NR-52310 Resources) or VSV-G (a gift from Tannishtha Reya (Addgene plasmid # 14888 ; http://n2t.net/addgene:14888 ; Addgene_14888) as control. detected: Addgene_36083detected: Addgene_12260detected: Addgene_14888Results 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 Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).
Results from JetFighter: We did not find any issues relating to colormaps.
About SciScore
SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore is not a substitute for expert review. SciScore checks for the presence and correctness of RRIDs (research resource identifiers) in the manuscript, and detects sentences that appear to be missing RRIDs. SciScore also checks to make sure that rigor criteria are addressed by authors. It does this by detecting sentences that discuss criteria such as blinding or power analysis. SciScore does not guarantee that the rigor criteria that it detects are appropriate for the particular study. Instead it assists authors, editors, and reviewers by drawing attention to sections of the manuscript that contain or should contain various rigor criteria and key resources. For details on the results shown here, including references cited, please follow this link.
-
SciScore for 10.1101/2020.05.21.20108951: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Written informed consent was obtained from the individuals recruited for this study . Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources In summary, pLV-S pseudotyped virus provides a valid screening tool for the presence of anti SARS-CoV-2 specific neutralizing antibodies in convalescent patient serum. anti SARS-CoV-2suggested: (Abcam Cat# ab273074, AB_2847846)Experimental Models: Cell Lines Sentences Resources , Vero E6 and 293T cells and compared … SciScore for 10.1101/2020.05.21.20108951: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Written informed consent was obtained from the individuals recruited for this study . Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources In summary, pLV-S pseudotyped virus provides a valid screening tool for the presence of anti SARS-CoV-2 specific neutralizing antibodies in convalescent patient serum. anti SARS-CoV-2suggested: (Abcam Cat# ab273074, AB_2847846)Experimental Models: Cell Lines Sentences Resources , Vero E6 and 293T cells and compared them to 293T cells transfected with plasmid encoding hACE2 receptor . Vero E6suggested: CVCL_XD71<div style="margin-bottom:8px"> <div><b>293T</b></div> <div>suggested: None</div> </div> </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">While untransfected 293T cells were half as efficient as hACE2 transfected cells , the MDCK , VeroE6 and A549 showed only a low level of transduction ( Fig 2) .</td><td style="min-width:100px;border-bottom:1px solid lightgray"> <div style="margin-bottom:8px"> <div><b>A549</b></div> <div>suggested: NCI-DTP Cat# A549, <a href="https://scicrunch.org/resources/Any/search?q=CVCL_0023">CVCL_0023</a></div> </div> </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">HEK293T were transfected with the following plasmids: 10 µg pLNCX-GFP , 1.2 µg pCMV-tat-HIV , pJK3 , and 3 µg of pL-VSV-G ( control ) or pCAGGS-S ( SARS-CoV-2) .</td><td style="min-width:100px;border-bottom:1px solid lightgray"> <div style="margin-bottom:8px"> <div><b>HEK293T</b></div> <div>suggested: KCB Cat# KCB 200744YJ, <a href="https://scicrunch.org/resources/Any/search?q=CVCL_0063">CVCL_0063</a></div> </div> </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">( E ) Transduction efficiency of pLV pseudotyped with CoV-2 Spike glycoprotein in HEK293T , Madin-Darby Canine Kidney cells ( MDCK)</td><td style="min-width:100px;border-bottom:1px solid lightgray"> <div style="margin-bottom:8px"> <div><b>MDCK</b></div> <div>suggested: CLS Cat# 602280/p823_MDCK_(NBL-2), <a href="https://scicrunch.org/resources/Any/search?q=CVCL_0422">CVCL_0422</a></div> </div> </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">), adenocarcinomic human alveolar ar basal epithelial cells (A549), and Vero (VeroE6) cells depicted as a percentage of efficiency in 293T cells transfected with plasmid encoding hACE2 receptor (hACE2293T).</td><td style="min-width:100px;border-bottom:1px solid lightgray"> <div style="margin-bottom:8px"> <div><b>Vero</b></div> <div>suggested: None</div> </div> </td></tr><tr><td style="min-width:100px;text-align:center; padding-top:4px;" colspan="2"><b>Recombinant DNA</b></td></tr><tr><td style="min-width:100px;text=align:center"><i>Sentences</i></td><td style="min-width:100px;text-align:center"><i>Resources</i></td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">293T cells were transfected with hACE2 plasmid ( a gift from Hyeryun Choe ( Addgene plasmid # 1786 ; http://n2t.net/addgene:1786 ; <a href="https://scicrunch.org/resources/Any/search?q=Addgene_1786">Addgene_1786</a> ) to produce hACE2-293T cells .</td><td style="min-width:100px;border-bottom:1px solid lightgray"> <div style="margin-bottom:8px"> <div><b></b></div> <div>suggested: <a href="https://scicrunch.org/resources/Any/search?q=Addgene_1786">Addgene_1786</a></div> </div> </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Generation of pseudotype particles: HEK293T cells ( 2 x 106 ) were plated in a 100-mm tissue culture dish and transfected the next day when they were about 75 % confluent w combination of the following plasmids: 9 µg of pLV-eGFP ( a gift from Pantelis Tsoulfas ( Addgene plasmid # 36083 ; http://n2t.net/addgene:36083 ; <a href="https://scicrunch.org/resources/Any/search?q=Addgene_36083">Addgene_36083</a>) , 9 µ psPAX2 ( a gift from Didier Trono ( Addgene plasmid # 12260 ; http://n2t.net/addgene:12 <a href="https://scicrunch.org/resources/Any/search?q=Addgene_12260">Addgene_12260</a>) , and 3 µg of pCAGGS-S ( SARS-CoV-2) ( Catalog No. NR-52310 Resources ) or VSV-G ( a gift from Tannishtha Reya ( Addgene plasmid # 14888 ; http://n2t.net/addgene:14888 ; <a href="https://scicrunch.org/resources/Any/search?q=Addgene_14888">Addgene_14888</a> ) as control .</td><td style="min-width:100px;border-bottom:1px solid lightgray"> <div style="margin-bottom:8px"> <div><b></b></div> <div>suggested: <a href="https://scicrunch.org/resources/Any/search?q=Addgene_36083">Addgene_36083</a></div> </div> <div style="margin-bottom:8px"> <div><b></b></div> <div>suggested: <a href="https://scicrunch.org/resources/Any/search?q=Addgene_12260">Addgene_12260</a></div> </div> <div style="margin-bottom:8px"> <div><b></b></div> <div>suggested: <a href="https://scicrunch.org/resources/Any/search?q=Addgene_14888">Addgene_14888</a></div> </div> </td></tr><tr><td style="min-width:100px;text-align:center; padding-top:4px;" colspan="2"><b>Software and Algorithms</b></td></tr><tr><td style="min-width:100px;text=align:center"><i>Sentences</i></td><td style="min-width:100px;text-align:center"><i>Resources</i></td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">The biological, chemical, and serological studies of the recently emerged SARS-CoV-2 will be greatly aided by the development and optimization of a suitable pseudotyping system.</td><td style="min-width:100px;border-bottom:1px solid lightgray"> <div style="margin-bottom:8px"> <div><b>SARS-CoV-2</b></div> <div>suggested: (Sino Biological Cat# 40143-R019, <a href="https://scicrunch.org/resources/Any/search?q=AB_2827973">AB_2827973</a>)</div> </div> </td></tr></table>Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).
Results from OddPub: We did not find a statement about open data. We also did not find a statement about open code. Researchers are encouraged to share open data when possible (see Nature blog).
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