Rapid response flow cytometric assay for the detection of antibody responses to SARS-CoV-2

Lapuente, Dennis; Maier, Clara; Irrgang, Pascal; Huebner, Julian; Peter, Sophia Antonia; Hoffmann, Markus; Ensser, Armin; Ziegler, Katharina; Winkler, Thomas H.; Birkholz, Thorsten; Kremer, Andreas E.; Steininger, Philipp; Korn, Klaus; Neipel, Frank; Ueberla, Klaus; Tenbusch, Matthias

SARS-CoV-2 has emerged as a previously unknown zoonotic coronavirus that spread worldwide causing a serious pandemic. While reliable nucleic acid-based diagnostic assays were rapidly available, there exists only a limited number of validated serological assays. Here, we evaluated a novel flow cytometric approach based on antigen-expressing HEK 293T cells to assess spike-specific IgG and IgM antibody responses. Analyses of 201 pre-COVID-19 sera proved a high assay specificity in comparison to commercially available CLIA and ELISA systems, while also revealing the highest sensitivity in specimens from PCR-confirmed SARS-CoV-2 infected patients. Additionally, a soluble Angiotensin-Converting-Enzyme 2 (ACE-2) variant was established as external standard to quantify spike-specific antibody responses on different assay platforms. In conclusion, our newly established flow cytometric assay allows sensitive and quantitative detection of SARS-CoV-2-specific antibodies, which can be easily adopted in different laboratories and does not rely on external supply of assay kits.