{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,19]],"date-time":"2026-02-19T05:25:31Z","timestamp":1771478731978,"version":"3.50.1"},"reference-count":42,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2024,1,25]],"date-time":"2024-01-25T00:00:00Z","timestamp":1706140800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,1,25]],"date-time":"2024-01-25T00:00:00Z","timestamp":1706140800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2021YFD1800100"],"award-info":[{"award-number":["2021YFD1800100"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100010038","name":"Earmarked Fund for China Agriculture Research System","doi-asserted-by":"publisher","award":["CARS-35"],"award-info":[{"award-number":["CARS-35"]}],"id":[{"id":"10.13039\/501100010038","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Appl Microbiol Biotechnol"],"published-print":{"date-parts":[[2024,12]]},"abstract":"<jats:sec>\n            <jats:title>Abstract<\/jats:title>\n            <jats:p>Pyroptosis is a newly discovered type of pro-inflammatory programmed cell death that plays a vital role in various processes such as inflammations, immune responses, and pathogen infections. As one of the main executioners of pyroptosis, gasdermin D (GSDMD) is a membrane pore-forming protein that typically exists in a self-inhibitory state. Once activated, GSDMD will be cleaved into an N-terminal fragment with pore-forming activity, becoming the key indicator of pyroptosis activation, and a C-terminal fragment. Although commercial antibodies against human and murine GSDMD proteins are currently available, their reactivity with porcine GSDMD (pGSDMD) is poor, which limits research on the biological functions of pGSDMD and pyroptosis in pigs in vivo and in vitro. Here, five monoclonal antibodies (mAbs) were prepared by immunizing BALB\/c mice with procaryotically expressed full-length pGSDMD, all of which did not cross react with human and murine GSDMD proteins. Epitope mapping demonstrated that 15H6 recognizes amino acids (aa) at positions 28\u201334 of pGSDMD (LQTSDRF), 19H3 recognizes 257\u2013260aa (PPQF), 23H10 and 27A10 recognize 78\u201382aa (GPFYF), and 25E2 recognizes 429\u2013435aa (PPTLLGS). The affinity constant and isotype of 15H6, 19H3, 23H10, 27A10, and 25E2 mAbs were determined to be 1.32\u2009\u00d7\u200910<jats:sup>\u22129<\/jats:sup>, 3.66\u2009\u00d7\u200910<jats:sup>\u22129<\/jats:sup>, 9.04\u2009\u00d7\u200910<jats:sup>\u22129<\/jats:sup>, 1.83\u2009\u00d7\u200910<jats:sup>\u22129<\/jats:sup>, and 8.00\u2009\u00d7\u200910<jats:sup>\u22128<\/jats:sup> mol\/L and IgG1\/\u03ba, IgG2a\/\u03ba, IgG2a\/\u03ba, IgG1\/\u03ba, and IgG1\/\u03ba, respectively. Heavy- and light-chain variable regions sequencing showed that the heavy-chain complementarity-determining region (CDR) sequences of all five mAbs are completely different, while the light-chain CDR sequences of the four mAbs that recognize the N-terminus of pGSDMD are identical. Our prepared mAbs provide valuable materials for studying pGSDMD function and pyroptosis.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Key points<\/jats:title>\n            <jats:p>\n              <jats:italic>\u2022 A total of five mouse anti-pGSDMD mAbs were prepared, of which four recognize the N-terminus of pGSDMD and one recognize its C-terminus.<\/jats:italic>\n            <\/jats:p>\n            <jats:p>\n              <jats:italic>\u2022 The main performance parameters of the five mAbs, including epitope, antibody titer, affinity constant, isotype, and heavy- and light-chain CDR, were characterized.<\/jats:italic>\n            <\/jats:p>\n            <jats:p>\n              <jats:italic>\u2022 All five mAbs specifically recognize pGSDMD protein and do not cross react with human and murine GSDMD proteins.<\/jats:italic>\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1007\/s00253-023-12938-x","type":"journal-article","created":{"date-parts":[[2024,1,25]],"date-time":"2024-01-25T03:02:31Z","timestamp":1706151751000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Preparation and characterization of monoclonal antibodies against porcine gasdermin D protein"],"prefix":"10.1007","volume":"108","author":[{"given":"Minhui","family":"Yang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xinna","family":"Ge","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lei","family":"Zhou","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xin","family":"Guo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jun","family":"Han","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0134-2761","authenticated-orcid":false,"given":"Yongning","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hanchun","family":"Yang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,1,25]]},"reference":[{"issue":"5","key":"12938_CR1","doi-asserted-by":"publisher","first-page":"1106","DOI":"10.1038\/s41423-020-00630-3","volume":"18","author":"D Bertheloot","year":"2021","unstructured":"Bertheloot D, Latz E, Franklin BS (2021) Necroptosis, pyroptosis and apoptosis: an intricate game of cell death. Cell Mol Immunol 18(5):1106\u20131121. https:\/\/doi.org\/10.1038\/s41423-020-00630-3","journal-title":"Cell Mol Immunol"},{"issue":"3","key":"12938_CR2","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1038\/s41577-019-0228-2","volume":"20","author":"P Broz","year":"2020","unstructured":"Broz P, Pelegr\u00edn P, Shao F (2020) The gasdermins, a protein family executing cell death and inflammation. Nat Rev Immunol 20(3):143\u2013157. https:\/\/doi.org\/10.1038\/s41577-019-0228-2","journal-title":"Nat Rev Immunol"},{"issue":"5","key":"12938_CR3","doi-asserted-by":"publisher","first-page":"1101","DOI":"10.1046\/j.1365-2958.1996.471410","volume":"21","author":"LM Chen","year":"1996","unstructured":"Chen LM, Kaniga K, Gal\u00e1n JE (1996) Salmonella spp. are cytotoxic for cultured macrophages. 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