Monkeypox virus H3L protein as the target antigen for developing neutralizing antibody and serological assay

Abstract The large number of atypical monkeypox (Mpox) cases caused by emerging monkeypox virus (MPXV) strains was recently found in countries and regions where the Mpox was not reported before. Diagnostic tools and therapeutic agents are important countermeasures for preventing Mpox outbreak. H3L p...

Full description

Saved in:
Bibliographic Details
Main Authors: I-Hsiang Huang, Guan-Chun Lai, Tai-Ling Chao, Wang-Da Liu, Sui-Yuan Chang, Shih-Chung Chang
Format: Article
Language:English
Published: Springer 2025-04-01
Series:Applied Microbiology and Biotechnology
Subjects:
Online Access:https://doi.org/10.1007/s00253-025-13466-6
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849237706370973696
author I-Hsiang Huang
Guan-Chun Lai
Tai-Ling Chao
Wang-Da Liu
Sui-Yuan Chang
Shih-Chung Chang
author_facet I-Hsiang Huang
Guan-Chun Lai
Tai-Ling Chao
Wang-Da Liu
Sui-Yuan Chang
Shih-Chung Chang
author_sort I-Hsiang Huang
collection DOAJ
description Abstract The large number of atypical monkeypox (Mpox) cases caused by emerging monkeypox virus (MPXV) strains was recently found in countries and regions where the Mpox was not reported before. Diagnostic tools and therapeutic agents are important countermeasures for preventing Mpox outbreak. H3L protein is the important surface antigen of MPXV for binding to host cell receptors and mediating viral infection. A broad range of murine anti-MPXV H3L monoclonal antibodies (mAbs) recognizing various binding epitopes have been generated in the study. The rapid test composed of the mAbs 4-2A and 3-3F can specifically detect H3L protein and MPXV virion. The mAb 3-3F exhibited strong MPXV neutralizing activity in a complement-dependent manner. Notably, 3-3F binds to a unique epitope within residues 35–89 of H3L protein. The serum samples collected from Mpox patients barely bound to the N-terminal portion of H3L protein ranging from 2 to 89 residues, indicating that the content of the 3-3F-like antibody is very low in Mpox patient sera. In contrast, the seropositivity was mostly observed using the C-terminal portion of H3L protein ranging from 185 to 282 residues as the target antigen in the immunoblot analysis. Taken together, the anti-MPXV H3L mAb can be developed as the Mpox diagnostic and therapeutic agents. Furthermore, H3L protein is the promising biomarker for serological analysis. Key Points •Anti-H3L mAbs can cross-react with H3L proteins in MPXV and VACV virions. •The LFIA rapid test using the mAbs 4-2A and 3-3F can specifically detect MPXV. •MPXV was neutralized by mAb 3-3F in a complement-dependent manner
format Article
id doaj-art-ac3f3a90d1454ae8a063b8b4334084cf
institution Kabale University
issn 1432-0614
language English
publishDate 2025-04-01
publisher Springer
record_format Article
series Applied Microbiology and Biotechnology
spelling doaj-art-ac3f3a90d1454ae8a063b8b4334084cf2025-08-20T04:01:52ZengSpringerApplied Microbiology and Biotechnology1432-06142025-04-01109111210.1007/s00253-025-13466-6Monkeypox virus H3L protein as the target antigen for developing neutralizing antibody and serological assayI-Hsiang Huang0Guan-Chun Lai1Tai-Ling Chao2Wang-Da Liu3Sui-Yuan Chang4Shih-Chung Chang5Department of Biochemical Science and Technology, College of Life Science, National Taiwan UniversityDepartment of Biochemical Science and Technology, College of Life Science, National Taiwan UniversityDepartment of Clinical Laboratory Sciences and Medical Biotechnology, College of Medicine, National Taiwan UniversityDepartment of Internal Medicine, National Taiwan University Hospital, College of Medicine, National Taiwan UniversityDepartment of Clinical Laboratory Sciences and Medical Biotechnology, College of Medicine, National Taiwan UniversityDepartment of Biochemical Science and Technology, College of Life Science, National Taiwan UniversityAbstract The large number of atypical monkeypox (Mpox) cases caused by emerging monkeypox virus (MPXV) strains was recently found in countries and regions where the Mpox was not reported before. Diagnostic tools and therapeutic agents are important countermeasures for preventing Mpox outbreak. H3L protein is the important surface antigen of MPXV for binding to host cell receptors and mediating viral infection. A broad range of murine anti-MPXV H3L monoclonal antibodies (mAbs) recognizing various binding epitopes have been generated in the study. The rapid test composed of the mAbs 4-2A and 3-3F can specifically detect H3L protein and MPXV virion. The mAb 3-3F exhibited strong MPXV neutralizing activity in a complement-dependent manner. Notably, 3-3F binds to a unique epitope within residues 35–89 of H3L protein. The serum samples collected from Mpox patients barely bound to the N-terminal portion of H3L protein ranging from 2 to 89 residues, indicating that the content of the 3-3F-like antibody is very low in Mpox patient sera. In contrast, the seropositivity was mostly observed using the C-terminal portion of H3L protein ranging from 185 to 282 residues as the target antigen in the immunoblot analysis. Taken together, the anti-MPXV H3L mAb can be developed as the Mpox diagnostic and therapeutic agents. Furthermore, H3L protein is the promising biomarker for serological analysis. Key Points •Anti-H3L mAbs can cross-react with H3L proteins in MPXV and VACV virions. •The LFIA rapid test using the mAbs 4-2A and 3-3F can specifically detect MPXV. •MPXV was neutralized by mAb 3-3F in a complement-dependent mannerhttps://doi.org/10.1007/s00253-025-13466-6Monkeypox virus (MPXV)H3L proteinLateral flow immunochromatographic assayNeutralizing antibodyComplementSerological assay
spellingShingle I-Hsiang Huang
Guan-Chun Lai
Tai-Ling Chao
Wang-Da Liu
Sui-Yuan Chang
Shih-Chung Chang
Monkeypox virus H3L protein as the target antigen for developing neutralizing antibody and serological assay
Applied Microbiology and Biotechnology
Monkeypox virus (MPXV)
H3L protein
Lateral flow immunochromatographic assay
Neutralizing antibody
Complement
Serological assay
title Monkeypox virus H3L protein as the target antigen for developing neutralizing antibody and serological assay
title_full Monkeypox virus H3L protein as the target antigen for developing neutralizing antibody and serological assay
title_fullStr Monkeypox virus H3L protein as the target antigen for developing neutralizing antibody and serological assay
title_full_unstemmed Monkeypox virus H3L protein as the target antigen for developing neutralizing antibody and serological assay
title_short Monkeypox virus H3L protein as the target antigen for developing neutralizing antibody and serological assay
title_sort monkeypox virus h3l protein as the target antigen for developing neutralizing antibody and serological assay
topic Monkeypox virus (MPXV)
H3L protein
Lateral flow immunochromatographic assay
Neutralizing antibody
Complement
Serological assay
url https://doi.org/10.1007/s00253-025-13466-6
work_keys_str_mv AT ihsianghuang monkeypoxvirush3lproteinasthetargetantigenfordevelopingneutralizingantibodyandserologicalassay
AT guanchunlai monkeypoxvirush3lproteinasthetargetantigenfordevelopingneutralizingantibodyandserologicalassay
AT tailingchao monkeypoxvirush3lproteinasthetargetantigenfordevelopingneutralizingantibodyandserologicalassay
AT wangdaliu monkeypoxvirush3lproteinasthetargetantigenfordevelopingneutralizingantibodyandserologicalassay
AT suiyuanchang monkeypoxvirush3lproteinasthetargetantigenfordevelopingneutralizingantibodyandserologicalassay
AT shihchungchang monkeypoxvirush3lproteinasthetargetantigenfordevelopingneutralizingantibodyandserologicalassay