Streif, Simon and Neckermann, Patrick and Spitzenberg, Clemens and Weiss, Katharina and Hoecherl, Kilian and Kulikowski, Kacper and Hahner, Sonja and Noelting, Christina and Einhauser, Sebastian and Peterhoff, David and Asam, Claudia and Wagner, Ralf and Baeumner, Antje J. J. (2023) Liposome-based high-throughput and point-of-care assays toward the quick, simple, and sensitive detection of neutralizing antibodies against SARS-CoV-2 in patient sera. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 415 (8). pp. 1421-1435. ISSN 1618-2642, 1618-2650
Full text not available from this repository. (Request a copy)Abstract
The emergence of severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) in 2019 caused an increased interest in neutralizing antibody tests to determine the immune status of the population. Standard live-virus-based neutralization assays such as plaque-reduction assays or pseudovirus neutralization tests cannot be adapted to the point-of-care (POC). Accordingly, tests quantifying competitive binding inhibition of the angiotensin-converting enzyme 2 (ACE2) receptor to the receptor-binding domain (RBD) of SARS-CoV-2 by neutralizing antibodies have been developed. Here, we present a new platform using sulforhodamine B encapsulating liposomes decorated with RBD as foundation for the development of both a fluorescent, highly feasible high-throughput (HTS) and a POC-ready neutralizing antibody assay. RBD-conjugated liposomes are incubated with serum and subsequently immobilized in an ACE2-coated plate or mixed with biotinylated ACE2 and used in test strip with streptavidin test line, respectively. Polyclonal neutralizing human antibodies were shown to cause complete binding inhibition, while S309 and CR3022 human monoclonal antibodies only caused partial inhibition, proving the functionality of the assay. Both formats, the HTS and POC assay, were then tested using 20 sera containing varying titers of neutralizing antibodies, and a control panel of sera including prepandemic sera and reconvalescent sera from respiratory infections other than SARS-CoV-2. Both assays correlated well with a standard pseudovirus neutralization test (r = 0.847 for HTS and r = 0.614 for POC format). Furthermore, excellent correlation (r = 0.868) between HTS and POC formats was observed. The flexibility afforded by liposomes as signaling agents using different dyes and sizes can hence be utilized in the future for a broad range of multianalyte neutralizing antibody diagnostics.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | ; Liposomes; Neutralizing antibodies; Point-of-care diagnostics; High-throughput screening; SARS-CoV-2 |
| Subjects: | 500 Science > 540 Chemistry & allied sciences |
| Divisions: | Medicine > Lehrstuhl für Medizinische Mikrobiologie und Hygiene Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik > Chemo- und Biosensorik (Prof. Antje J. Bäumner, formerly Prof. Wolfbeis) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 09 Mar 2024 09:22 |
| Last Modified: | 09 Mar 2024 09:22 |
| URI: | https://pred.uni-regensburg.de/id/eprint/59274 |
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