Impedance‐based monitoring of titration and neutralization assays with VSV‐G and SARS‐CoV‐2‐spike pseudoviruses

• DOI: 10.1002/appl.202400097
• Publication Date: 2024-08-28T05:30:02Z
• Authors: Anne‐Kathrin Mildner; Sebastian Einhauser; Stefanie Michaelis; Klara Rogalla v. ...; Ralf Wagner; Joachim Wegener
• Keywords: spike protein; 2019-20 coronavirus outbreak; Coronavirus

Abstract Since cell‐based virus neutralization assays are still the gold standard to assess a patient's immune protection against given virus, they of utmost importance for serodiagnosis, convalescent plasma therapy, and vaccine development. Monitoring emergence characteristics neutralizing antibodies in an outbreak situation, confirming as correlates from infection testing vaccine‐induced potency antibody responses, quests automated, fast, parallel assays. We developed impedance‐based sensor platform (electric cell‐substrate impedance sensing, ECIS) providing time‐resolved monitoring host cell response viral pseudotypes. For validation, assay was compared with state‐of‐the‐art quantification virus‐induced reporter protein expression independent indicator neutralization. Vesicular stomatitis (VSV) derived pseudoviruses encoding green fluorescent (GFP) autologous G (VSV‐G) initial binding membrane were used HEK293T both, optical readout. Virus‐induced cytopathic effects (CPE) detectable low pseudotype concentrations (multiplicity 1) profiles soon 5–10 h after concentration‐dependent manner. Neutralization efficacy α‐VSV‐G determined time courses IC 50 values favorably fluorescence measurements virus‐borne GFP expression. Sera COVID‐19 patients tested successfully SARS‐CoV‐2 by incubating VSV, pseudotyped spike protein, different sera before exposure recordings. In summary: (i) ECIS applied detect virus‐mediated neutralization; (ii) Impedance‐based allows reducing h; (iii) is easily adapted other virus‐based diseases scalable high‐throughput.