Acetylcholinesterase (AChE) is a key enzyme in the nervous system capable of breaking down acetylcholine (ACh). ACh is a critical neurotransmitter that signals between nerve cells and muscles. Organophosphates (OPs), used as insecticides and nerve agents, irreversibly inhibit AChE by forming a covalent bond with the enzyme’s active site. Since exposure to these nerve agents can quickly lead to death, many researchers have strived to sdiscover new drugs that will reverse the inhibition of AChE by such agents. The goal was to develop a high-throughput miniaturized assay for testing large numbers of compounds for AChE reactivation quickly and reliably. Here, we describe our miniaturized assay that utilises the sarin surrogate nitrophenyl isopropyl methylphosphonate (NIMP) in an Amplex Red coupled assay and its validation with 42 compounds selected from an in silico virtual screen.
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Published on: Sep 20, 2025 Pages: 1-8
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DOI: 10.17352/ab.000011
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