Development of a rapid image-based high-content imaging screening assay to evaluate therapeutic antibodies against the monkeypox virus.

Autor: Kota KP; Tonix Pharmaceuticals Inc, Frederick, MD, USA., Ziółkowska NE; Tonix Pharmaceuticals Inc, Frederick, MD, USA., Wei J; Tonix Pharmaceuticals Inc, Frederick, MD, USA., Peng J; Tonix Pharmaceuticals Inc, Frederick, MD, USA., Ordonez D; Tonix Pharmaceuticals Inc, Frederick, MD, USA., Raney C; Tonix Pharmaceuticals Inc, Frederick, MD, USA., Prigge J; Bioqual Inc., Rockville, MD, USA., Hooper JW; United States Army Medical Institute of Infectious Diseases, Frederick, MD, USA., Awasthi M; Tonix Pharmaceuticals Inc, Frederick, MD, USA., Goebel SJ; Tonix Pharmaceuticals Inc, Frederick, MD, USA., Zabel BA; Curia Global Inc., Albany, NY, USA., Nasar F; Tonix Pharmaceuticals Inc, Frederick, MD, USA., Lederman S; Tonix Pharmaceuticals Inc, Frederick, MD, USA., Bavari S; Tonix Pharmaceuticals Inc, Frederick, MD, USA. Electronic address: sina.bavari@tonixpharma.com.
Jazyk: angličtina
Zdroj: Antiviral research [Antiviral Res] 2023 Feb; Vol. 210, pp. 105513. Date of Electronic Publication: 2022 Dec 30.
DOI: 10.1016/j.antiviral.2022.105513
Abstrakt: Antibody-based therapy is emerging as a critical therapeutic countermeasure to treat acute viral infections by offering rapid protection against clinical disease. The advancements in structural biology made it feasible to rationalize monoclonal antibodies (mAbs) by identifying key and, possibly, neutralizing epitopes of viral proteins for therapeutic purposes. A critical component in assessing mAbs during pandemics requires the development of rapid but detailed methods to detect and quantitate the neutralization activity. In this study, we developed and optimized two high-content image (HCI)-based assays: one to detect viral proteins by staining and the second to quantify cytopathic viral effects by a label-free phenotypic assay. These assays were employed to screen for therapeutic antibodies against the monkeypox virus (MPXV) using surrogate poxviruses such as vaccinia virus (VACV). Plaque-based neutralization results confirmed the HCI data. The phenotypic assay found pox virus-induced syncytia formation in various cells, and we were able to quantitate and use this phenotype to screen mAbs. The HCI identified several potent VACV-neutralizing antibodies that showed in vitro efficacy against both clades of MPXV. In addition, a combination study of ST-246/tecovirimat/TPOXX a single neutralizing antibody Ab-40, showed synergistic activity against VACV in an in-vitro neutralization assay. This rapid high-content method utilizing state-of-the-art technologies enabled the evaluation of hundreds of mAbs quickly to identify several potent anti-MPXV neutralizing mAbs for further development.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2022 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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