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The mutant strains of Mpox virus (MPXV) were found with higher infectivity and greater capability for sustained human-to-human transmission, posing a significant public health threat. MPXV A29L, a protein homolog of vaccinia virus (VACV) A27L, plays an important role in viral attachment to host cell membranes. Therefore, MPXV A29L is considered the diagnostic target and the potential vaccine candidate for eliciting neutralizing antibodies and protective immune responses. In response to the escalating Mpox outbreak, three monoclonal antibodies (mAbs) (2-9B, 3-8G, and 2-5H) targeting the different domains of MPXV A29L have been developed in the study. Among them, 2-5H is highly specific for MPXV A29L without exhibiting cross-reactivity with VACV A27L. The antibody pairing composed of 2-5H and 3-8G has been developed as the lateral flow immunochromatographic assay for specific detection of MPXV A29L. However, these three mAbs were unable to inhibit A29L binding to heparin column or prevent MPXV infection in the neutralization test assays. The results of the serological assays using the truncated A29L fragments as the antigens showed that the Mpox patient sera contained significantly lower levels of antibodies targeting the N-terminal 1\u201334 residues of A29L, suggesting that the N-terminal portion of A29L is less immunogenic upon natural infection.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Key points<\/jats:title>\n            <jats:p>\n              <jats:italic>\u2022 MAbs 2-9B, 3-8G, and 2-5H neither interrupted A29L binding to heparin nor neutralized MPXV.<\/jats:italic>\n            <\/jats:p>\n            <jats:p>\n              <jats:italic>\u2022 The LFIA composed of 3-8G and 2-5H can specifically distinguish MPXV A29L from VACV A27L.<\/jats:italic>\n            <\/jats:p>\n            <jats:p>\n              <jats:italic>\u2022 Mpox patient sera contained lower levels of antibodies targeting the N-terminal portion of A29L.<\/jats:italic>\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1007\/s00253-024-13361-6","type":"journal-article","created":{"date-parts":[[2024,11,21]],"date-time":"2024-11-21T06:47:19Z","timestamp":1732171639000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Monkeypox virus A29L protein as the target for specific diagnosis and serological analysis"],"prefix":"10.1007","volume":"108","author":[{"given":"Chia-Yu","family":"Liang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tai-Ling","family":"Chao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chong\u2010Syun","family":"Chao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wang-Da","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu-Chen","family":"Cheng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sui-Yuan","family":"Chang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5015-8178","authenticated-orcid":false,"given":"Shih-Chung","family":"Chang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,11,21]]},"reference":[{"issue":"23","key":"13361_CR1","doi-asserted-by":"crossref","first-page":"12355","DOI":"10.1128\/JVI.01593-09","volume":"83","author":"MR Benhnia","year":"2009","unstructured":"Benhnia MR, McCausland MM, Laudenslager J, Granger SW, Rickert S, Koriazova L, Tahara T, Kubo RT, Kato S, Crotty S (2009a) Heavily isotype-dependent protective activities of human antibodies against vaccinia virus extracellular virion antigen B5. 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