Rapid, high throughput, automated detection of SARS-CoV-2 neutralizing antibodies against native-like vaccine and delta variant spike trimers
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Abstract
Traditional cellular and live-virus methods for detection of SARS-CoV-2 neutralizing antibodies (nAbs) are labor- and time-intensive, and thus not suited for routine use in the clinical lab to predict vaccine efficacy and natural immune protection. Here, we report the development and validation of a rapid, high throughput method for measuring SARS-CoV-2 nAbs against native-like trimeric spike proteins. This assay uses a blockade of hACE-2 binding (BoAb) approach in an automated digital immunoassay on the Quanterix HD-X platform. BoAb assays using vaccine and delta variant viral strains showed strong correlation with cell-based pseudovirus and live-virus neutralization activity. Importantly, we were able to detect similar patterns of delta variant resistance to neutralization in samples with paired vaccine and delta variant BoAb measurements. Finally, we screened clinical samples from patients with or without evidence of SARS-CoV-2 exposure by a single-dilution screening version of our assays, finding significant nAb activity only in exposed individuals. In principle, these assays offer a rapid, robust, and scalable alternative to time-, skill-, and cost-intensive standard methods for measuring SARS-CoV-2 nAb levels.
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SciScore for 10.1101/2022.02.01.22270279: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics Consent: Samples: Samples were sourced from various studies with IRB 00001663 at Emory University after obtaining the approval and consent from Institutional Review Board and samples (n=300) are tested on The Quanterix HD-X which uses single molecule arrays (SIMOAS) of femtoliter-sized reaction chambers etched into a disk to detect single enzyme labeled proteins.
IRB: Samples: Samples were sourced from various studies with IRB 00001663 at Emory University after obtaining the approval and consent from Institutional Review Board and samples (n=300) are tested on The Quanterix HD-X which uses single molecule arrays (SIMOAS) of femtoliter-sized reaction chambers etched into a disk to detect …SciScore for 10.1101/2022.02.01.22270279: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics Consent: Samples: Samples were sourced from various studies with IRB 00001663 at Emory University after obtaining the approval and consent from Institutional Review Board and samples (n=300) are tested on The Quanterix HD-X which uses single molecule arrays (SIMOAS) of femtoliter-sized reaction chambers etched into a disk to detect single enzyme labeled proteins.
IRB: Samples: Samples were sourced from various studies with IRB 00001663 at Emory University after obtaining the approval and consent from Institutional Review Board and samples (n=300) are tested on The Quanterix HD-X which uses single molecule arrays (SIMOAS) of femtoliter-sized reaction chambers etched into a disk to detect single enzyme labeled proteins.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
Experimental Models: Cell Lines Sentences Resources Briefly, to produce SARS-CoV-2 WT and Delta pseudoviruses, an expression plasmid bearing codon optimized SARS-CoV-2 full-length S plasmid (parental sequence Wuhan-1, Genbank #: MN908947.3) was co-transfected into HEK293T cells (ATCC#CRL-11268) with plasmids encoding non-surface proteins for lentivirus production and a lentiviral backbone plasmid expressing a Luciferase-IRES-ZsGreen reporter, HIV-1 Tat and Rev packing plasmids (BEI Resources) and pseudoviruses harvested after 48 hours of post transfection and performed titration. HEK293Tsuggested: ATCC Cat# CRL-11268, RRID:CVCL_1926)Pseudoviruses were mixed with serial dilutions of plasma or antibodies and then added to monolayers of ACE-2-overexpressing 293T cells (BEI Resources), in duplicate. 293Tsuggested: KCB Cat# KCB 200744YJ, RRID:CVCL_0063)Software and Algorithms Sentences Resources Statistical Analysis: GraphPad PRISM version 9 was used to perform the statistical analysis. GraphPad PRISMsuggested: (GraphPad Prism, RRID:SCR_002798)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.
Results from scite Reference Check: We found no unreliable references.
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