Transmembrane Protease Serine 2 (TMPRSS2) Inhibitors Screened from an Fv-antibody Library for Preventing SARS-CoV-2 Infection

Overview

Journal RSC Med Chem

Publisher Royal Society of Chemistry

Specialty Chemistry

Date 2025 Feb 24

PMID 39990164

Authors

Jaeyong Jung

Jeong Soo Sung

Soonil Kwon

Hyung Eun Bae

Min-Jung Kang

Joachim Jose

Misu Lee

Jae-Chul Pyun

Affiliations

Soon will be listed here.

Abstract

Fv-antibodies targeting the transmembrane protease serine 2 (TMPRSS2) were screened from an Fv-antibody library for inhibiting SARS-CoV-2 infection. Fv-antibodies were derived from the variable region of heavy-chain immunoglobulin G (IgG), which consisted of three complementarity-determining regions (CDRs) and frame regions (FRs). The Fv-antibody library was prepared through site-directed mutagenesis of CDR3 region. The proteolytic cleavage site (S2' site) of TMPRSS2 on the spike protein (SP) of SARS-CoV-2 was used as a screening probe for the library. Two Fv-antibodies were screened and subsequently expressed as soluble recombinant proteins. The binding affinities of the expressed Fv-antibodies were estimated using a surface plasmon resonance (SPR) biosensor. The two expressed Fv-antibodies specifically bound to the active site of TMPRSS2 which interacts with S2' site in the proprotein convertase (PPC) region. The neutralizing activities of the two expressed Fv-antibodies were demonstrated using a cell-based infection assay with pseudo-viruses that expressed the SP of four types of SARS-CoV-2 variants: Wu-1 (D614), Delta (B.1.617.2), Omicron (BA.2), and Omicron (BA.4/5). Additionally, a docking simulation was performed to analyze the interaction between the screened Fv-antibodies and the active sites of TMPRSS2.

References

Wrapp D, Wang N, Corbett K, Goldsmith J, Hsieh C, Abiona O . Cryo-EM structure of the 2019-nCoV spike in the prefusion conformation. Science. 2020; 367(6483):1260-1263. PMC: 7164637. DOI: 10.1126/science.abb2507. View

Kim T, Park J, Jung J, Sung J, Kwon S, Bae H . A one-step immunoassay based on switching peptides for diagnosis of porcine epidemic diarrhea virus (PEDV) using screened Fv-antibodies. J Mater Chem B. 2024; 12(15):3751-3763. DOI: 10.1039/d4tb00066h. View

Kumar S, Bose D, Suryawanshi H, Sabharwal H, Mapa K, Maiti S . Specificity of RSG-1.2 peptide binding to RRE-IIB RNA element of HIV-1 over Rev peptide is mainly enthalpic in origin. PLoS One. 2011; 6(8):e23300. PMC: 3154333. DOI: 10.1371/journal.pone.0023300. View

Walls A, Park Y, Tortorici M, Wall A, McGuire A, Veesler D . Structure, Function, and Antigenicity of the SARS-CoV-2 Spike Glycoprotein. Cell. 2020; 181(2):281-292.e6. PMC: 7102599. DOI: 10.1016/j.cell.2020.02.058. View

Jung J, Bong J, Sung J, Lee S, Lee M, Kang M . Fluorescein and Rhodamine B-Binding Domains from Autodisplayed Fv-Antibody Library. Bioconjug Chem. 2021; 32(10):2213-2223. DOI: 10.1021/acs.bioconjchem.1c00376. View

Jung J, Kwon S, Sung J, Bae H, Kang M, Jose J . Screened Fv-Antibodies against the Angiotensin-Converting Enzyme 2 (ACE2) Receptor Neutralizing the Infection of SARS-CoV-2. ACS Pharmacol Transl Sci. 2024; 7(12):3914-3920. PMC: 11651164. DOI: 10.1021/acsptsci.4c00441. View

Chen M, Zhang X . Construction and applications of SARS-CoV-2 pseudoviruses: a mini review. Int J Biol Sci. 2021; 17(6):1574-1580. PMC: 8071765. DOI: 10.7150/ijbs.59184. View

Jung J, Bong J, Lee S, Kim M, Sung J, Lee M . Screening of Fv Antibodies with Specific Binding Activities to Monosodium Urate and Calcium Pyrophosphate Dihydrate Crystals for the Diagnosis of Gout and Pseudogout. ACS Appl Bio Mater. 2022; 4(4):3388-3397. DOI: 10.1021/acsabm.0c01680. View

Fraser B, Beldar S, Seitova A, Hutchinson A, Mannar D, Li Y . Structure and activity of human TMPRSS2 protease implicated in SARS-CoV-2 activation. Nat Chem Biol. 2022; 18(9):963-971. DOI: 10.1038/s41589-022-01059-7. View

10.

Sung J, Bong J, Lee S, Jung J, Kang M, Lee M . One-step immunoassay for food allergens based on screened mimotopes from autodisplayed F-antibody library. Biosens Bioelectron. 2022; 202:113976. DOI: 10.1016/j.bios.2022.113976. View

11.

Xu J, Davis M . Diversity in the CDR3 region of V(H) is sufficient for most antibody specificities. Immunity. 2000; 13(1):37-45. DOI: 10.1016/s1074-7613(00)00006-6. View

12.

Lan J, Ge J, Yu J, Shan S, Zhou H, Fan S . Structure of the SARS-CoV-2 spike receptor-binding domain bound to the ACE2 receptor. Nature. 2020; 581(7807):215-220. DOI: 10.1038/s41586-020-2180-5. View

13.

Hristova S, Zhivkov A . Omicron Coronavirus: pH-Dependent Electrostatic Potential and Energy of Association of Spike Protein to ACE2 Receptor. Viruses. 2023; 15(8). PMC: 10460073. DOI: 10.3390/v15081752. View

14.

McCallum M, Park Y, Stewart C, Sprouse K, Addetia A, Brown J . Human coronavirus HKU1 recognition of the TMPRSS2 host receptor. Cell. 2024; 187(16):4231-4245.e13. DOI: 10.1016/j.cell.2024.06.006. View

15.

DAngelo S, Ferrara F, Naranjo L, Erasmus M, Hraber P, Bradbury A . Many Routes to an Antibody Heavy-Chain CDR3: Necessary, Yet Insufficient, for Specific Binding. Front Immunol. 2018; 9:395. PMC: 5852061. DOI: 10.3389/fimmu.2018.00395. View

16.

Zahradnik J, Marciano S, Shemesh M, Zoler E, Harari D, Chiaravalli J . SARS-CoV-2 variant prediction and antiviral drug design are enabled by RBD in vitro evolution. Nat Microbiol. 2021; 6(9):1188-1198. DOI: 10.1038/s41564-021-00954-4. View

17.

Pedelacq J, Cabantous S, Tran T, Terwilliger T, Waldo G . Engineering and characterization of a superfolder green fluorescent protein. Nat Biotechnol. 2005; 24(1):79-88. DOI: 10.1038/nbt1172. View

18.

Qi X, Ke B, Feng Q, Yang D, Lian Q, Li Z . Construction and immunogenic studies of a mFc fusion receptor binding domain (RBD) of spike protein as a subunit vaccine against SARS-CoV-2 infection. Chem Commun (Camb). 2020; 56(61):8683-8686. DOI: 10.1039/d0cc03263h. View

19.

Salleh M, Deris Z . Molecular Characterization of Human TMPRSS2 Protease Polymorphic Variants and Associated SARS-CoV-2 Susceptibility. Life (Basel). 2022; 12(2). PMC: 8876804. DOI: 10.3390/life12020231. View

20.

Yoo G, Saenger T, Bong J, Jose J, Kang M, Pyun J . Co-autodisplay of Z-domains and bovine caseins on the outer membrane of E. coli. Biochim Biophys Acta. 2015; 1848(12):3126-33. DOI: 10.1016/j.bbamem.2015.09.018. View