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SARS-CoV-2 Mpro Assay Kit: A Key Tool for Evaluating Inhibitors and Understanding Mutations
Olgotrelvir, a dual inhibitor of SARS-CoV-2 Mpro and cathepsin L, as a standalone antiviral oral intervention candidate for COVID-19
SARS-CoV-2 Mpro Assay Kit: A Key Tool for Evaluating Inhibitors and Understanding Mutations
Introduction
Although the SARS-CoV-2 pandemic is firmly in the world’s rear-view mirror, its variants are still giving people reasons to stay home. The latest variants, known collectively as FLiRT, include KP.2, JN.1.7, and other emerging KP and JN subvariants. These variants are characterized by critical mutations in positions 456, 346, and 572 of the spike protein, which can potentially enhance viral binding affinity and immune evasion. Specifically, the mutation at position 456 is linked to increased transmissibility, while mutations at 346 and 572 may contribute to antibody resistance. As a result of ongoing SARS-CoV-2 research and monitoring efforts, Aurora BioLabs continues to provide high-quality SARS-CoV-2 products, such as assay kits and reagents, to support scientists in tracking and combating these evolving variants.
Link to Research Article: Olgotrelvir Research on Cell
Mechanism of Action: Dual Inhibition of Mpro and Cathepsin L
Olgotrelvir differentiates itself from conventional antivirals by targeting two key components essential for SARS-CoV-2 replication and entry:
SARS-CoV-2 Main Protease (Mpro): Mpro is crucial for viral replication as it processes the viral polyproteins at specific cleavage sites. Inhibition of Mpro halts viral replication, making it a prime target for antiviral drug development.
Cathepsin L: Cathepsin L is a host protease that facilitates viral entry by priming the spike protein. By inhibiting Cathepsin L, Olgotrelvir effectively blocks the virus’ ability to enter host cells.
The dual inhibition mechanism broadens the spectrum of Olgotrelvir’s antiviral activity, making it a promising candidate for both prophylactic and therapeutic applications.
SARS-CoV-2 Mpro, also referred to as SARS-CoV-2 Main protease and SARS-CoV-2 3CL protease, plays an essential role in viral replication by processing the polyproteins 1ab at 11 distinct cleavage sites. This protease is highly conserved among coronaviruses, making it an ideal target for broad-spectrum antiviral drugs. Mpro operates through a cysteine-histidine catalytic dyad mechanism, where its active site cleaves the polyproteins to release non-structural proteins essential for viral replication and assembly. Inhibiting the activity of this enzyme effectively halts viral replication by preventing the formation of functional viral replication complexes. Therefore, Mpro is a highly promising target for the development of anti-coronaviral therapeutic agents, including both small-molecule inhibitors and peptide-based drugs.
Aurora SARS-CoV-2 Mpro assay kit is a homogeneous FRET-based assay specifically designed for high-throughput screening of Mpro inhibitors. This assay stands out for its speed, convenience, and reliability, requiring only 30 minutes and two simple steps to obtain results. In the first step, the recombinant Mpro enzyme is preincubated with potential inhibitor compounds for 30 minutes, allowing sufficient interaction time. The reaction is then initiated by adding the substrate solution in the second step. The fluorescent intensity, which correlates with Mpro activity, is measured using a fluorescent plate reader at excitation wavelengths of 340-360 nm and emission wavelengths of 460-480 nm.
In the featured study, the Aurora SARS-CoV-2 Mpro assay kit played a crucial role in evaluating the dual inhibitory activity of Olgotrelvir against both Mpro and cathepsin L. By providing a rapid and precise means to assess the inhibitory effects of candidate compounds, the assay facilitated the identification of Olgotrelvir as a promising oral antiviral intervention. The kit's high sensitivity and reproducibility were instrumental in generating reliable data, which ultimately contributed to advancing antiviral research.
Aurora SARS-CoV-2 Mpro Assay Kit: Supporting Antiviral Research
Aurora Biolabs offers a SARS-CoV-2 Mpro assay kit, designed to facilitate the screening of Mpro inhibitors like Olgotrelvir. This homogeneous FRET-based assay provides a fast and reliable platform for evaluating potential antivirals.
Assay Features:
Requires only 30 minutes and two steps for completion.
Compatible with fluorescent plate readers measuring at excitation wavelengths of 340-360nm and emission wavelengths of 460-480nm.
The kit is ideal for research labs focused on developing and testing novel SARS-CoV-2 inhibitors.
Kit Contents
2x Assay Buffer (20ml)
0.5M DTT (200µl)
0.5 mM Substrate (10µl)
Recombinant SARS-CoV-2 Mpro (5µg)
One 96-well plate
Materials Needed (Not Supplied)
A microplate reader capable of measuring fluorescence at excitation wavelengths of 340-360nm and emission wavelengths of 460-480nm
Aurora Biolabs: Innovation in Antiviral Research
Aurora Biolabs is a leading provider of high-quality research tools and assay kits designed to support scientists in their quest for groundbreaking discoveries. With a commitment to innovation and scientific excellence, Aurora Biolabs offers a diverse range of products tailored to the needs of researchers working on SARS-CoV-2 and other viral pathogens. Our expertise in assay development and recombinant protein production ensures that our products meet the highest standards of reliability and reproducibility.
Related Products
BHE20700015: SARS-CoV-2 Mpro (3CL Protease) Assay Kit
BHP20700065: Recombinant SARS-CoV-2 Mpro, 3CL protease
BHP20700067: Recombinant SARS-CoV-2 Papain-like Protease (PLpro, NSP3), CF
BHP20700066: Recombinant SARS-CoV-2 Helicase (NSP13)
Conclusion
The research on Olgotrelvir represents a significant advancement in the fight against COVID-19. Its dual inhibition mechanism offers a novel approach to antiviral therapy, addressing both viral replication and entry. Aurora Biolabs remains committed to supporting such groundbreaking research by providing high-quality assay kits and related products for SARS-CoV-2 research. In addition, Biohippo Inc. plays a crucial role as a trusted partner in the life sciences field, offering a comprehensive range of innovative research products and services. By collaborating with leading suppliers and leveraging a robust e-commerce platform, Biohippo ensures that researchers have access to cutting-edge tools and solutions to accelerate scientific discovery.
Reference
Hu Q, Xiong Y, Zhu GH, Zhang YN, Zhang YW, Huang P, Ge GB. The SARS-CoV-2 main protease (Mpro): Structure, function, and emerging therapies for COVID-19. MedComm (2020). 2022 Jul 14;3(3):e151. doi: 10.1002/mco2.151. PMID: 35845352; PMCID: PMC9283855.
