1. Academic Validation
  2. High-Throughput cell-based immunofluorescence assays against influenza

High-Throughput cell-based immunofluorescence assays against influenza

  • SLAS Discov. 2024 Jan;29(1):66-76. doi: 10.1016/j.slasd.2023.10.008.
Yohanka Martinez-Gzegozewska 1 Lynn Rasmussen 2 Sara McKellip 2 Anna Manuvakhova 2 N Miranda Nebane 2 Andrew J Reece 2 Pedro Ruiz 2 Melinda Sosa 2 Robert Bostwick 2 Paige Vinson 2
Affiliations

Affiliations

  • 1 Scientific Platforms Division, Southern Research, High-Throughput Screening Center, Birmingham, Alabama, United States. Electronic address: ymartinez@southernresearch.org.
  • 2 Scientific Platforms Division, Southern Research, High-Throughput Screening Center, Birmingham, Alabama, United States.
Abstract

A rapid drug discovery response to influenza outbreaks with the potential to reach pandemic status could help minimize the virus's impact by reducing the time to identify anti-influenza drugs. Although several anti-influenza strategies have been considered in the search for new drugs, only a few therapeutic agents are approved for clinical use. The cytopathic effect induced by the Influenza Virus in Madin Darby canine kidney (MDCK) cells has been widely used for high-throughput anti-influenza drug screening, but the fact that the MDCK cells are not human cells constitutes a disadvantage when searching for new therapeutic agents for human use. We have developed a highly sensitive cell-based imaging assay for the identification of inhibitors of influenza A and B virus that is high-throughput compatible using the A549 human cell line. The assay has also been optimized for the assessment of the neutralizing effect of anti-influenza Antibodies in the absence of trypsin, which allows testing of purified Antibodies and serum samples. This assay platform can be applied to full high-throughput screening campaigns or later stages requiring quantitative potency determinations for structure-activity relationships.

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