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Optimized pipeline for generating highly potent neutralizing Affitins: Application to SARS-CoV-2 spike protein
Thomas Maurice1, Mélanie Laporte2, Nicolas Barquilla2
1Nantes Université, Univ Angers, INSERM, CNRS, Immunology and New Concepts in ImmunoTherapy, INCIT, UMR 1302/EMR6001, F-44000, Nantes, France; Nantes Université, CHU Nantes, CNRS, Inserm, BioCore, US16, Plateforme P2R, SFR Bonamy, F-44000, Nantes, France.
Artificial proteins called Affitins show strong potential as antiviral therapies against SARS-CoV-2. Engineered Affitin multimers neutralize the virus effectively, offering a promising alternative to traditional antibody treatments.
Area of Science:
- Biotechnology
- Virology
- Protein Engineering
Background:
- Viral infections pose a significant global health burden, necessitating novel therapeutic strategies.
- Protein-based therapies, like monoclonal antibodies (mAbs), offer high efficacy but can be costly and complex.
- Affitins, small and stable artificial proteins, present a promising alternative scaffold for antiviral drug development.
Purpose of the Study:
- To engineer and evaluate Affitin-based antiviral agents targeting the SARS-CoV-2 spike protein.
- To explore different multimerization formats of Affitins for enhanced neutralization potency.
- To assess the efficacy and stability of Affitin constructs against SARS-CoV-2 variants.
Main Methods:
- Utilized ribosome display technology to generate diverse Affitin libraries.
- Engineered Affitins into multimeric formats, including dimers, trimers, and novel hexamers.
- Quantified neutralization activity using IC₅₀ values and assessed binding affinity and thermal stability.
Main Results:
- Engineered Affitin dimers and trimers demonstrated potent SARS-CoV-2 neutralization across variants (IC₅₀ as low as 39 pM).
- Achieved high affinity (22.8 pM) and thermal stability (84.7 °C) with multimeric Affitin constructs.
- Developed the first hexameric Affitins, exhibiting exceptional neutralization efficacy (IC₅₀ = 0.8 pM) against the B.1.351 variant.
Conclusions:
- Affitins represent a versatile and potent scaffold for developing next-generation antiviral therapies.
- Multimeric Affitin constructs offer advantages in potency and size compared to traditional antibodies.
- These findings expand the therapeutic arsenal against emerging viral threats like SARS-CoV-2.
