The Use of Heterologous Antigens for Biopanning Enables the Selection of Broadly Neutralizing Nanobodies Against SARS-CoV-2 | Sciact - CRIS-system of Boreskov Institute of Catalysis

The Use of Heterologous Antigens for Biopanning Enables the Selection of Broadly Neutralizing Nanobodies Against SARS-CoV-2 Full article

Journal

Antibodies
ISSN: 2073-4468

Output data

Year: 2025, Volume: 14, Number: 1, Article number : 23, Pages count : 16 DOI: 10.3390/antib14010023

Tags

nanobodies; single-domain antibodies; VHH; recombinant antibodies; SARS-CoV-2; phage library; affinity selection; biopanning; AlphaFold; protein–protein docking; molecular dynamics

Authors

Aripov Vazirbek S. ¹ , Zaykovskaya Anna V. ¹ , Mechetina Ludmila V. ² , Najakshin Alexander M. ² , Bondar Alexander A. ³ , Arkhipov Sergey G. ⁴ , Mustaev Egor A. ^5,4 , Ilyina Margarita G. ⁴ , Borisevich Sophia S. ⁴ , Ilyichev Alexander A. ¹ , Nesmeyanova Valentina S. ¹ , Isaeva Anastasia A. ¹ , Volosnikova Ekaterina A. ¹ , Shcherbakov Dmitry N. ¹ , Volkova Natalia V. ¹

Affiliations

1	State Research Center of Virology and Biotechnology VECTOR, Rospotrebnadzor, Koltsovo 630559, Russia
2	Institute of Molecular and Cellular Biology, Siberian Branch of Russian Academy of Sciences, Novosibirsk 630090, Russia
3	Genomics Core Facility, Institute of Chemical Biology and Fundamental Medicine Siberian Branch of Russian Academy of Sciences, Novosibirsk 630090, Russia
4	Synchrotron Radiation Facility—Siberian Circular Photon Source “SKlF” Boreskov Institute of Catalysis of Siberian Branch of the Russian Academy of Sciences, Koltsovo 630559, Russia
5	Department of Natural Sciences, Novosibirsk State University, Novosibirsk 630090, Russia

Background: Since the emergence of SARS-CoV-2 in the human population, the virus genome has undergone numerous mutations, enabling it to enhance transmissibility and evade acquired immunity. As a result of these mutations, most monoclonal neutralizing antibodies have lost their efficacy, as they are unable to neutralize new variants. Antibodies that neutralize a broad range of SARS-CoV-2 variants are of significant value in combating both current and potential future variants, making the identification and development of such antibodies an ongoing critical goal. This study discusses the strategy of using heterologous antigens in biopanning rounds. Methods: After four rounds of biopanning, nanobody variants were selected from a phage display library. Immunochemical methods were used to evaluate their specificity to the S protein of various SARS-CoV-2 variants, as well as to determine their competitive ability against ACE2. Viral neutralization activity was analyzed. A three-dimensional model of nanobody interaction with RBD was constructed. Results: Four nanobodies were obtained that specifically bind to the receptor-binding domain (RBD) of the SARS-CoV-2 spike glycoprotein and exhibit neutralizing activity against various SARS-CoV-2 strains. Conclusions: The study demonstrates that performing several rounds of biopanning with heterologous antigens allows the selection of nanobodies with a broad reactivity spectrum. However, the fourth round of biopanning does not lead to the identification of nanobodies with improved characteristics.

Aripov V.S. , Zaykovskaya A.V. , Mechetina L.V. , Najakshin A.M. , Bondar A.A. , Arkhipov S.G. , Mustaev E.A. , Ilyina M.G. , Borisevich S.S. , Ilyichev A.A. , Nesmeyanova V.S. , Isaeva A.A. , Volosnikova E.A. , Shcherbakov D.N. , Volkova N.V.
The Use of Heterologous Antigens for Biopanning Enables the Selection of Broadly Neutralizing Nanobodies Against SARS-CoV-2
Antibodies. 2025. V.14. N1. 23 :1-16. DOI: 10.3390/antib14010023 Scopus OpenAlex publication_identifier_short.sciact_skif_identifier_type

Submitted:	Jan 29, 2025
Published print:	Mar 1, 2025
Accepted:	Mar 5, 2025
Published online:	Mar 7, 2025

Scopus:	2-s2.0-105001105859
OpenAlex:	W4408226623
publication_identifier.sciact_skif_identifier_type:	3962

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