Rapid and sensitive detection of SARS-CoV-2 antibodies by biolayer interferometry

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1. Version published to 10.1038/s41598-020-78895-x

   Dec 10, 2020
2. 

   ScreenIT

   Jul 20, 2020

   SciScore for 10.1101/2020.07.17.20156281: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: First, human serum samples (n = 25) collected in 2017 were from study participants enrolled in an Institutional Review Board-approved study for development of Lyme disease and other diagnostic tests.
   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
   Reagents and supplies: Phosphate buffered saline (PBS) tablets (Sigma P4417), Tween-20 (Fisher BP337), dry milk powder (RPI 50488786), ELISA plates (Corning 3590), Goat anti-Human IgG Fc HRP (Thermo Fisher A18817), OPD tablets (Pierce PI34006), bovine serum albumin …

   SciScore for 10.1101/2020.07.17.20156281: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: First, human serum samples (n = 25) collected in 2017 were from study participants enrolled in an Institutional Review Board-approved study for development of Lyme disease and other diagnostic tests.
   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
   Reagents and supplies: Phosphate buffered saline (PBS) tablets (Sigma P4417), Tween-20 (Fisher BP337), dry milk powder (RPI 50488786), ELISA plates (Corning 3590), Goat anti-Human IgG Fc HRP (Thermo Fisher A18817), OPD tablets (Pierce PI34006), bovine serum albumin (BSA) (Fisher BP1600), ChonBlock (Chondrex 9068), Biotinylated SARS-CoV-2 protein RBD His AviTag (Acro Biosystems SPD-C82E9), 4 nm Colloidal Gold-AffiPure Goat Anti-Human IgG Fcg fragment specific (Jackson ImmunoResearch 109-185-098), 4 nm Colloidal Gold-AffiPure Goat Anti-Human Serum IgA alpha chain specific (Jackson ImmunoResearch 109-185-011), Human coronavirus spike glycoprotein Antibody, Rabbit PAb, Antigen Affinity Purified (Sino Biological 40021-T60), SARS-CoV-2 (2019-nCoV) spike Antibody, Rabbit PAb, Antigen Affinity Purified (Sino Biological 40589-T62), SARS-CoV-2 (2019-nCoV) spike Antibody, Rabbit MAb (40150-R007), Anti-SARS-CoV S Therapeutic Antibody (CR3022) (Creative Biolabs MRO-1214LC), Octet Anti-Penta-His (HIS1K) sensor tips (Sartorius ForteBio 18-5120), Octet Streptavidin (SA) sensor tips (Sartorius ForteBio 18-5109), tilted bottom (TW384) microplates (Sartorius ForteBio 18-5080), electroporation cuvettes (MaxCyte SOC4), suspension adapted CHO-S cells (Thermo Fisher R80007).	anti-Human IgG suggested: (Thermo Fisher Scientific Cat# A18817, RRID:AB_2535594) Anti-Human Serum IgA suggested: (Jackson ImmunoResearch Labs Cat# 109-185-011, RRID:AB_2337743) Human coronavirus spike glycoprotein Antibody, Rabbit PAb, Antigen Affinity Purified suggested: None Anti-SARS-CoV S suggested: None CR3022 suggested: (Imported from the IEDB Cat# CR3022, RRID:AB_2848080) Anti-Penta-His (HIS1K suggested: None
   The human monoclonal antibody CR3022 antibody, which is reactive to the RBD of both SARS-CoV-1 and SARS-CoV-2, was used as a positive control 34-36.	SARS-CoV-2 suggested: None
   The assay plate was prepared as follows: column 1 (BLI assay buffer), column 2 (2-12 μg/ml RBD or prefusion Spike in BLI assay buffer), column 3 (25% ChonBlock in BLI assay buffer), column 4 (plasma/serum samples diluted 1:8 in 25% ChonBlock in BLI assay buffer), column 5 (BLI assay buffer), and column 6 (4 nm Colloidal Gold-AffiPure Goat Anti-Human IgG or IgA secondary antibody diluted 1:10 in BLI assay buffer).	IgA suggested: None
   Baseline1 (60 sec) in column 1 (Equilibration), Loading (120 sec or 600 sec) in column 2 (Antigen Loading: RBD or prefusion Spike, respectively), Baseline2 (60 sec) in column 3 (Wash), Association1 (600 sec) in column 4 (Total Antibody Binding), Baseline3 (60 sec) in column 5 (Wash), and Association2 (180 sec) in column 6 (Detection: anti-human IgG or IgA).	Baseline2 suggested: None Association1 suggested: None Baseline3 suggested: None Association2 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   For protein production, CHO-S cells were transfected with purified endotoxin-free DNA using flow electroporation technology (MaxCyte).	CHO-S suggested: RRID:CVCL_7183)
   Software and Algorithms
   Sentences	Resources
   For purification of RBD-biotin, media was supplemented with 20 mM Tris pH 8.0 and 150 mM NaCl (TBS), 1 mM EDTA, and BioLock and loaded onto a StrepTrap column.	BioLock suggested: None
   The AUC values were calculated by fitting a four-parameter logistic regression model to the OD490 data of each sample using the curve fit algorithm from the SciPy Python Library 42 followed by integration between the upper and lower bounds of the data.	SciPy suggested: (SciPy, RRID:SCR_008058)
   This Python program named bli_plotter is available for adaption to other BLI-ISA studies and has been deposited on GitHub (https://github.com/nlorigroach/bli_plotter).	Python suggested: (IPython, RRID:SCR_001658)

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   Results from OddPub: Thank you for sharing your code and data.

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   Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:

   We do acknowledge a caveat of our BLI-ISA method is the need for BLI instrumentation, however this instrument is becoming more commonplace at research institutions, and in many cases the acquisition of such an instrument would be beneficial due to reduced labor, faster data acquisition time, and reduced data variability compared to other methods. We also acknowledge the relatively small numbers of pre-pandemic samples used for this proof-of-principal study are not sufficient to precisely define the lower limit of detection, however evaluation of more pre-pandemic samples will ultimately make this limit clear. In a broader sense, BLI-ISA can be adapted, multiplexed, and performed in a high-throughput fashion. Straightforward adaptions can allow for measurement of antibodies against different antigens, as we demonstrated for RBD and prefusion Spike, and/or detection of different antibody isotypes, as we demonstrated for IgG and IgA antibodies. There is also potential to measure antibodies in other biological specimens (e.g. breastmilk, saliva) and also measure other clinically relevant, non-antibody biological molecules in human and animal specimens. Finally, BLI-ISA has the potential to be performed in a high-throughput fashion. While many institutions have BLI instruments that measure 8 or 16 biosensors at a time, the Octet HTX instrument can measure 96 biosensors at a time. In addition, we found that RBD-biotin could be pre-loaded onto SA biosensors with no loss in signal ov...

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   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).

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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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3. 

   Version published to 10.1101/2020.07.17.20156281v1 on medRxiv

   Jul 19, 2020