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Related Concept Videos

Inhibitors of Viral Protein Synthesis01:30

Inhibitors of Viral Protein Synthesis

Protein synthesis is indispensable for viral replication, as viruses lack the cellular machinery required for this process and must hijack the host's translational apparatus. In response, host cells deploy a critical innate immune defense involving interferons, specialized cytokines that play a central role in inhibiting viral propagation.Upon viral detection, infected cells release interferons that bind to receptors on adjacent uninfected cells, activating the JAK-STAT signaling pathway and...

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Bio-layer Interferometry for Measuring Kinetics of Protein-protein Interactions and Allosteric Ligand Effects
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Characterization of antiviral compounds using Bio-Layer Interferometry.

Zachary C Lorson1,2, William M McFadden1,2, Grace Neilsen1,2

  • 1Center for ViroScience and Cure, Laboratory of Biochemical Pharmacology, Department of Pediatrics, Emory University School of Medicine, 1760 Haygood Drive NE, Atlanta, GA.

Biorxiv : the Preprint Server for Biology
|August 8, 2025
PubMed
Summary

Optimized biolayer interferometry (BLI) protocols effectively characterize small molecule-protein interactions for antiviral discovery. This high-throughput method overcomes common pitfalls, enabling precise affinity measurements for drug development.

Keywords:
Biolayer interferometry (BLI)biomolecular interactionscapsid protein (CA)human immunodeficiency virus (HIV)lenacapavir (LEN)main protease nsp5nirmatrelvir (NIR)severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)

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Area of Science:

  • Biochemistry and Molecular Biology
  • Drug Discovery and Development
  • Biophysical Chemistry

Background:

  • Small molecule-protein interactions are crucial for biological functions and disease treatment.
  • Biolayer interferometry (BLI) offers a sensitive, high-throughput approach for studying these interactions, particularly in antiviral discovery.
  • Challenges like nonspecific binding and ligand drift can hinder BLI accuracy.

Purpose of the Study:

  • To present optimized BLI protocols for characterizing small molecule-protein interactions.
  • To address and provide solutions for common BLI pitfalls.
  • To demonstrate BLI's utility in calculating binding affinities for clinically relevant antiviral compounds.

Main Methods:

  • Development and application of optimized BLI protocols.
  • Characterization of interactions between PF74 and HIV-1 capsid protein (CA).
  • Analysis of Lenacapavir (LEN) with HIV-1 CA and Nirmatrelvir (NIR) with SARS-CoV-2 Mpro.

Main Results:

  • Demonstrated BLI as a powerful tool for calculating binding affinities (KD values) from micro to sub-nanomolar ranges.
  • Quantified binding affinities for HIV-1 CA interactions with PF74 and LEN.
  • Confirmed the covalent interaction between Nirmatrelvir and SARS-CoV-2 Mpro.

Conclusions:

  • Optimized BLI protocols facilitate accurate characterization of small molecule-protein binding.
  • BLI is effective for determining binding affinities and interaction types for antiviral drug candidates.
  • These protocols support high-throughput screening and testing of novel antivirals and derivatives.