Research output: Contribution to journal › Article › peer-review
The Use of Heterologous Antigens for Biopanning Enables the Selection of Broadly Neutralizing Nanobodies Against SARS-CoV-2. / Aripov, Vazirbek S.; Zaykovskaya, Anna V.; Mechetina, Ludmila V. et al.
In: Antibodies, Vol. 14, No. 1, 23, 07.03.2025.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - The Use of Heterologous Antigens for Biopanning Enables the Selection of Broadly Neutralizing Nanobodies Against SARS-CoV-2
AU - Aripov, Vazirbek S.
AU - Zaykovskaya, Anna V.
AU - Mechetina, Ludmila V.
AU - Najakshin, Alexander M.
AU - Bondar, Alexander A.
AU - Arkhipov, Sergey G.
AU - Mustaev, Egor A.
AU - Ilyina, Margarita G.
AU - Borisevich, Sophia S.
AU - Ilyichev, Alexander A.
AU - Nesmeyanova, Valentina S.
AU - Isaeva, Anastasia A.
AU - Volosnikova, Ekaterina A.
AU - Shcherbakov, Dmitry N.
AU - Volkova, Natalia V.
N1 - The work was carried out as part of the state assignment 23/21 of the main activities plan of the State Research Center of Virology and Biotechnology ‘Vector’, Rospotrebnadzor, for 2025.
PY - 2025/3/7
Y1 - 2025/3/7
N2 - 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.
AB - 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.
KW - AlphaFold
KW - SARS-CoV-2
KW - VHH
KW - affinity selection
KW - biopanning
KW - molecular dynamics
KW - nanobodies
KW - phage library
KW - protein–protein docking
KW - recombinant antibodies
KW - single-domain antibodies
UR - https://www.mendeley.com/catalogue/5db466f4-9b3a-3608-8760-4a799c7ee30f/
UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-105001105859&origin=inward&txGid=5507249b00103de65413274b4c3063b8
U2 - 10.3390/antib14010023
DO - 10.3390/antib14010023
M3 - Article
C2 - 40136472
VL - 14
JO - Antibodies
JF - Antibodies
SN - 2073-4468
IS - 1
M1 - 23
ER -
ID: 65140166