{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T03:39:46Z","timestamp":1790739586126,"version":"4.1.0"},"reference-count":38,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2015,5,21]],"date-time":"2015-05-21T00:00:00Z","timestamp":1432166400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2015,5,21]],"date-time":"2015-05-21T00:00:00Z","timestamp":1432166400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Mesothelin is an emerging cell surface target in mesothelioma and other solid tumors. Most antibody drug candidates recognize highly immunogenic Region I (296\u2013390) on mesothelin. Here, we report a group of high-affinity non-Region I rabbit monoclonal antibodies. These antibodies do not compete for mesothelin binding with the immunotoxin SS1P that binds Region I of mesothelin. One pair of antibodies (YP218 and YP223) is suitable to detect soluble mesothelin in a sandwich ELISA with high sensitivity. The new assay can also be used to measure serum mesothelin concentration in mesothelioma patients, indicating its potential use for monitoring patients treated with current antibody therapies targeting Region I. The antibodies are highly specific and sensitive in immunostaining of mesothelioma. To explore their use in tumor therapy, we have generated the immunotoxins based on the Fv of these antibodies. One immunotoxin (YP218 Fv-PE38) exhibits potent anti-tumor cytotoxicity towards primary mesothelioma cell lines <jats:italic>in vitro<\/jats:italic> and an NCI-H226 xenograft tumor in mice. Furthermore, we have engineered a humanized YP218 Fv that retains full binding affinity for mesothelin-expressing cancer cells. In conclusion, with their unique binding properties, these antibodies may be promising candidates for monitoring and treating mesothelioma and other mesothelin-expressing cancers.<\/jats:p>","DOI":"10.1038\/srep09928","type":"journal-article","created":{"date-parts":[[2015,5,21]],"date-time":"2015-05-21T16:06:21Z","timestamp":1432224381000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":51,"title":["New High Affinity Monoclonal Antibodies Recognize Non-Overlapping Epitopes On Mesothelin For Monitoring And Treating Mesothelioma"],"prefix":"10.1038","volume":"5","author":[{"given":"Yi-Fan","family":"Zhang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yen","family":"Phung","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Gao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seiji","family":"Kawa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raffit","family":"Hassan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ira","family":"Pastan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mitchell","family":"Ho","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2015,5,21]]},"reference":[{"key":"BFsrep09928_CR1","doi-asserted-by":"publisher","first-page":"136","DOI":"10.1073\/pnas.93.1.136","volume":"93","author":"K Chang","year":"1996","unstructured":"Chang, K. & Pastan, I. Molecular cloning of mesothelin, a differentiation antigen present on mesothelium, mesotheliomas and ovarian cancers. Proc. Natl. Acad. Sci. U S A . 93, 136\u2013140 (1996).","journal-title":"Proc. Natl. Acad. Sci. U S A"},{"key":"BFsrep09928_CR2","doi-asserted-by":"publisher","first-page":"2907","DOI":"10.1158\/0008-5472.CAN-14-0337","volume":"74","author":"I Pastan","year":"2014","unstructured":"Pastan, I. & Hassan, R. Discovery of Mesothelin and Exploiting It as a Target for Immunotherapy. Cancer Res. 74, 2907\u20132912 (2014).","journal-title":"Cancer Res."},{"key":"BFsrep09928_CR3","doi-asserted-by":"publisher","first-page":"276","DOI":"10.2174\/1871520611313020014","volume":"13","author":"Z Tang","year":"2013","unstructured":"Tang, Z., Qian, M. & Ho, M. The role of mesothelin in tumor progression and targeted therapy. Anticancer Agents. Med. Chem. 13, 276\u2013280 (2013).","journal-title":"Anticancer Agents. Med. Chem."},{"key":"BFsrep09928_CR4","doi-asserted-by":"publisher","first-page":"46","DOI":"10.1016\/j.ejca.2007.08.028","volume":"44","author":"R Hassan","year":"2008","unstructured":"Hassan, R. & Ho, M. Mesothelin targeted cancer immunotherapy. Eur. J. Cancer 44, 46\u201353 (2008).","journal-title":"Eur. J. Cancer"},{"key":"BFsrep09928_CR5","doi-asserted-by":"publisher","first-page":"1541","DOI":"10.1200\/JCO.2011.39.6671","volume":"30","author":"K Hollevoet","year":"2012","unstructured":"Hollevoet, K. et al. Serum Mesothelin for Diagnosing Malignant Pleural Mesothelioma: An Individual Patient Data Meta-Analysis. J. Clin. Oncol. 30, 1541\u20131549 (2012).","journal-title":"J. Clin. Oncol."},{"key":"BFsrep09928_CR6","doi-asserted-by":"publisher","first-page":"261","DOI":"10.2217\/bmm.11.18","volume":"5","author":"A Cristaudo","year":"2011","unstructured":"Cristaudo, A., Bonotti, A., Simonini, S., Bruno, R. & Foddis, R. Soluble markers for diagnosis of malignant pleural mesothelioma. Biomark Med. 5, 261\u2013273 (2011).","journal-title":"Biomark Med."},{"key":"BFsrep09928_CR7","doi-asserted-by":"publisher","first-page":"11531","DOI":"10.1073\/pnas.96.20.11531","volume":"96","author":"N Scholler","year":"1999","unstructured":"Scholler, N. et al. Soluble member(s) of the mesothelin\/megakaryocyte potentiating factor family are detectable in sera from patients with ovarian carcinoma. Proc. Natl. Acad. Sci. U S A . 96, 11531\u201311536 (1999).","journal-title":"Proc. Natl. Acad. Sci. U S A"},{"key":"BFsrep09928_CR8","doi-asserted-by":"publisher","first-page":"447","DOI":"10.1158\/1078-0432.CCR-05-1477","volume":"12","author":"R Hassan","year":"2006","unstructured":"Hassan, R. et al. Detection and Quantitation of Serum Mesothelin, a Tumor Marker for Patients with Mesothelioma and Ovarian Cancer. Clin Cancer Res. 12, 447\u2013453 (2006).","journal-title":"Clin Cancer Res."},{"key":"BFsrep09928_CR9","doi-asserted-by":"publisher","first-page":"1751","DOI":"10.1158\/1055-9965.EPI-06-0479","volume":"15","author":"M Ho","year":"2006","unstructured":"Ho, M. et al. Mesothelin Is Shed from Tumor Cells. Cancer Epidemiol. Biomarkers Prev. 15, 1751 (2006).","journal-title":"Cancer Epidemiol. Biomarkers Prev."},{"key":"BFsrep09928_CR10","doi-asserted-by":"publisher","first-page":"6132","DOI":"10.1158\/1078-0432.CCR-10-2275","volume":"16","author":"R Hassan","year":"2010","unstructured":"Hassan, R. et al. Phase I Clinical Trial of the Chimeric Anti-Mesothelin Monoclonal Antibody MORAb-009 in Patients with Mesothelin-Expressing Cancers. Clin. Cancer Res. 16, 6132\u20136138 (2010).","journal-title":"Clin. Cancer Res."},{"key":"BFsrep09928_CR11","doi-asserted-by":"publisher","first-page":"416","DOI":"10.1158\/1535-7163.MCT-12-0731","volume":"12","author":"Z Tang","year":"2013","unstructured":"Tang, Z. et al. A Human Single-Domain Antibody Elicits Potent Antitumor Activity by Targeting an Epitope in Mesothelin Close to the Cancer Cell Surface. Mol. Cancer Ther. 12, 416\u2013426 (2013).","journal-title":"Mol. Cancer Ther."},{"key":"BFsrep09928_CR12","doi-asserted-by":"publisher","first-page":"3937","DOI":"10.1158\/1078-0432.CCR-03-0801","volume":"10","author":"R Hassan","year":"2004","unstructured":"Hassan, R., Bera, T. & Pastan, I. Mesothelin: A New Target for Immunotherapy. Clin. Cancer Res. 10, 3937\u20133942 (2004).","journal-title":"Clin. Cancer Res."},{"key":"BFsrep09928_CR13","doi-asserted-by":"publisher","first-page":"2020","DOI":"10.1002\/ijc.25557","volume":"128","author":"M Ho","year":"2011","unstructured":"Ho, M., Feng, M., Fisher, R. J., Rader, C. & Pastan, I. A novel high-affinity human monoclonal antibody to mesothelin. Int. J. Cancer. 128, 2020\u20132030 (2011).","journal-title":"Int. J. Cancer."},{"key":"BFsrep09928_CR14","doi-asserted-by":"publisher","first-page":"208ra147","DOI":"10.1126\/scitranslmed.3006941","volume":"5","author":"R Hassan","year":"2013","unstructured":"Hassan, R. et al. Major Cancer Regressions in Mesothelioma After Treatment with an Anti-Mesothelin Immunotoxin and Immune Suppression. Sci. Transl. Med. 5, 208ra147 (2013).","journal-title":"Sci. Transl. Med."},{"key":"BFsrep09928_CR15","doi-asserted-by":"publisher","first-page":"3739","DOI":"10.1074\/jbc.M806776200","volume":"284","author":"O Kaneko","year":"2009","unstructured":"Kaneko, O. et al. A Binding Domain on Mesothelin for CA125\/MUC16. J. Biol. Chem. 284, 3739\u20133749 (2009).","journal-title":"J. Biol. Chem."},{"key":"BFsrep09928_CR16","doi-asserted-by":"publisher","first-page":"9190","DOI":"10.1074\/jbc.M312372200","volume":"279","author":"A Rump","year":"2004","unstructured":"Rump, A. et al. Binding of ovarian cancer antigen CA125\/MUC16 to mesothelin mediates cell adhesion. J Biol Chem . 279, 9190\u20139198 (2004).","journal-title":"J Biol Chem"},{"key":"BFsrep09928_CR17","doi-asserted-by":"publisher","first-page":"9348","DOI":"10.1073\/pnas.92.20.9348","volume":"92","author":"H Spieker-Polet","year":"1995","unstructured":"Spieker-Polet, H., Sethupathi, P., Yam, P. C. & Knight, K. L. Rabbit monoclonal antibodies: generating a fusion partner to produce rabbit-rabbit hybridomas. Proc. Natl. Acad. Sci. U S A . 92, 9348\u20139352 (1995).","journal-title":"Proc. Natl. Acad. Sci. U S A"},{"key":"BFsrep09928_CR18","doi-asserted-by":"publisher","first-page":"592","DOI":"10.4161\/mabs.20933","volume":"4","author":"Y Phung","year":"2012","unstructured":"Phung, Y., Gao, W., Man, Y.-G., Nagata, S. & Ho, M. High-affinity monoclonal antibodies to cell surface tumor antigen glypican-3 generated through a combination of peptide immunization and flow cytometry screening. mAbs. 4, 592\u2013599 (2012).","journal-title":"mAbs."},{"key":"BFsrep09928_CR19","first-page":"5151","volume":"32","author":"J Zhang","year":"2012","unstructured":"Zhang, J. et al. Loss of Mesothelin Expression by Mesothelioma Cells Grown In Vitro Determines Sensitivity to Anti-Mesothelin Immunotoxin SS1P. Anticancer Res. 32, 5151\u20135158 (2012).","journal-title":"Anticancer Res."},{"key":"BFsrep09928_CR20","unstructured":"Pastan, I. & Ho, M. in Antibody Engineering Vol. 2 (eds Roland Kontermann & Stefan Du\u00a8bel ) Ch. 10, 127\u2013146 (Springer-Verlag."},{"key":"BFsrep09928_CR21","doi-asserted-by":"publisher","first-page":"123","DOI":"10.7150\/jca.2.123","volume":"2","author":"M Feng","year":"2011","unstructured":"Feng, M., Zhang, J., Anver, M., Hassan, R. & Ho, M. In Vivo Imaging of Human Malignant Mesothelioma Grown Orthotopically in the Peritoneal Cavity of Nude Mice. J Cancer . 2, 123\u2013131 (2011).","journal-title":"J Cancer"},{"key":"BFsrep09928_CR22","doi-asserted-by":"publisher","first-page":"319","DOI":"10.1007\/978-3-642-01144-3_21","volume":"Ch. 21","author":"D Williams","year":"2010","unstructured":"Williams, D., Matthews, D. & Jones, T. in Antibody Engineering (eds Roland Kontermann & Stefan D\u00fcbel ) Ch. 21, 319\u2013339 Springer Berlin Heidelberg 2010).","journal-title":"Antibody Engineering"},{"key":"BFsrep09928_CR23","doi-asserted-by":"publisher","first-page":"1612","DOI":"10.1016\/S0140-6736(03)14794-0","volume":"362","author":"BWS Robinson","year":"2003","unstructured":"Robinson, B. W. S. et al. Mesothelin-family proteins and diagnosis of mesothelioma. The Lancet. 362, 1612\u20131616 (2003).","journal-title":"The Lancet."},{"key":"BFsrep09928_CR24","doi-asserted-by":"publisher","first-page":"5926","DOI":"10.1158\/1078-0432.CCR-11-1235","volume":"17","author":"X Du","year":"2011","unstructured":"Du, X., Xiang, L., Mackall, C. & Pastan, I. Killing of Resistant Cancer Cells with Low Bak by a Combination of an Antimesothelin Immunotoxin and a TRAIL Receptor 2 Agonist Antibody. Clin. Cancer Res. 17, 5926\u20135934 (2011).","journal-title":"Clin. Cancer Res."},{"key":"BFsrep09928_CR25","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1097\/CJI.0000000000000010","volume":"37","author":"K Hollevoet","year":"2014","unstructured":"Hollevoet, K., Antignani, A., FitzGerald, D. J. & Pastan, I. Combining the Antimesothelin Immunotoxin SS1P With the BH3-mimetic ABT-737 Induces Cell Death in SS1P-resistant Pancreatic Cancer Cells. J. Immunother. 37, 8\u201315 (2014).","journal-title":"J. Immunother."},{"key":"BFsrep09928_CR26","doi-asserted-by":"publisher","first-page":"727","DOI":"10.1016\/0161-5890(83)90050-0","volume":"20","author":"K PM","year":"1983","unstructured":"PM, K., R, G. & RD, P. Rabbit anti-carbohydrate antibody elicited by the lymphocyte mitogenic glycoprotein from Wistaria floribunda seeds. Mol. Immunol. 20, 727\u2013735 (1983).","journal-title":"Mol. Immunol."},{"key":"BFsrep09928_CR27","doi-asserted-by":"publisher","first-page":"325","DOI":"10.1016\/S0022-2836(02)01232-9","volume":"325","author":"M Popkov","year":"2003","unstructured":"Popkov, M. et al. Rabbit Immune Repertoires as Sources for Therapeutic Monoclonal Antibodies: The Impact of Kappa Allotype-correlated Variation in Cysteine Content on Antibody Libraries Selected by Phage Display. J. Mol. Biol. 325, 325\u2013335 (2003).","journal-title":"J. Mol. Biol."},{"key":"BFsrep09928_CR28","doi-asserted-by":"publisher","first-page":"13668","DOI":"10.1074\/jbc.275.18.13668","volume":"275","author":"C Rader","year":"2000","unstructured":"Rader, C. et al. The Rabbit Antibody Repertoire as a Novel Source for the Generation of Therapeutic Human Antibodies. J. Biol. Chem. 275, 13668\u201313676 (2000).","journal-title":"J. Biol. Chem."},{"key":"BFsrep09928_CR29","doi-asserted-by":"publisher","first-page":"e9072","DOI":"10.1371\/journal.pone.0009072","volume":"5","author":"Y Yu","year":"2010","unstructured":"Yu, Y. et al. A Humanized Anti-VEGF Rabbit Monoclonal Antibody Inhibits Angiogenesis and Blocks Tumor Growth in Xenograft Models. PloS. ONE. 5, e9072 (2010).","journal-title":"PloS. ONE."},{"key":"BFsrep09928_CR30","doi-asserted-by":"publisher","first-page":"5840","DOI":"10.1158\/1078-0432.CCR-05-0578","volume":"11","author":"M Onda","year":"2005","unstructured":"Onda, M. et al. New Monoclonal Antibodies to Mesothelin Useful for Immunohistochemistry, Fluorescence-Activated Cell Sorting, Western Blotting and ELISA. Clin. Cancer Res. 11, 5840\u20135846 (2005).","journal-title":"Clin. Cancer Res."},{"key":"BFsrep09928_CR31","doi-asserted-by":"publisher","first-page":"3814","DOI":"10.1158\/1078-0432.CCR-04-2304","volume":"11","author":"M Ho","year":"2005","unstructured":"Ho, M. et al. Humoral Immune Response to Mesothelin in Mesothelioma and Ovarian Cancer Patients. Clin. Cancer Res. 11, 3814\u20133820 (2005).","journal-title":"Clin. Cancer Res."},{"key":"BFsrep09928_CR32","doi-asserted-by":"publisher","first-page":"1520","DOI":"10.1158\/1055-9965.EPI-08-0039","volume":"17","author":"I Hellstrom","year":"2008","unstructured":"Hellstrom, I. et al. Anti-Mesothelin Antibodies and Circulating Mesothelin Relate to the Clinical State in Ovarian Cancer Patients. Cancer Epidemiol. Biomarkers Prev. 17, 1520\u20131526 (2008).","journal-title":"Cancer Epidemiol. Biomarkers Prev."},{"key":"BFsrep09928_CR33","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1016\/S0022-2836(02)01237-8","volume":"325","author":"S Ewert","year":"2003","unstructured":"Ewert, S., Huber, T., Honegger, A. & Pl\u00fcckthun, A. Biophysical Properties of Human Antibody Variable Domains. J. Mol. Biol. 325, 531\u2013553 (2003).","journal-title":"J. Mol. Biol."},{"key":"BFsrep09928_CR34","doi-asserted-by":"publisher","first-page":"2238","DOI":"10.1158\/1055-9965.EPI-10-0346","volume":"19","author":"J Creaney","year":"2010","unstructured":"Creaney, J. et al. Serum Mesothelin for Early Detection of Asbestos-Induced Cancer Malignant Mesothelioma. Cancer Epidemiol Biomarkers Prev. 19, 2238\u20132246 (2010).","journal-title":"Cancer Epidemiol Biomarkers Prev."},{"key":"BFsrep09928_CR35","doi-asserted-by":"publisher","first-page":"569","DOI":"10.1136\/thx.2006.068114","volume":"62","author":"J Creaney","year":"2007","unstructured":"Creaney, J. et al. Soluble mesothelin in effusions: a useful tool for the diagnosis of malignant mesothelioma. Thorax. 62, 569\u2013576 (2007).","journal-title":"Thorax."},{"key":"BFsrep09928_CR36","doi-asserted-by":"publisher","first-page":"666","DOI":"10.1373\/clinchem.2006.079327","volume":"53","author":"HL Beyer","year":"2007","unstructured":"Beyer, H. L. et al. MESOMARK\u2122: A Potential Test for Malignant Pleural Mesothelioma. Clinical. Chemist. 53, 666\u2013672 (2007).","journal-title":"Clinical. Chemist."},{"key":"BFsrep09928_CR37","doi-asserted-by":"publisher","first-page":"3311","DOI":"10.1002\/cncr.28875","volume":"120","author":"R Hassan","year":"2014","unstructured":"Hassan, R. et al. Phase 1 study of the antimesothelin immunotoxin SS1P in combination with pemetrexed and cisplatin for front-line therapy of pleural mesothelioma and correlation of tumor response with serum mesothelin, megakaryocyte potentiating factor and cancer antigen 125. Cancer . 120, 3311\u20133319 (2014).","journal-title":"Cancer"},{"key":"BFsrep09928_CR38","doi-asserted-by":"publisher","first-page":"4587","DOI":"10.1200\/JCO.2006.06.8999","volume":"24","author":"GL Ceresoli","year":"2006","unstructured":"Ceresoli, G. L. et al. Early Response Evaluation in Malignant Pleural Mesothelioma by Positron Emission Tomography With [18F]Fluorodeoxyglucose. J. Clin. Oncol. 24, 4587\u20134593 (2006).","journal-title":"J. Clin. Oncol."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/srep09928","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep09928.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep09928.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,1,5]],"date-time":"2023-01-05T13:12:19Z","timestamp":1672924339000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/srep09928"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,5,21]]},"references-count":38,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2015,9,22]]}},"alternative-id":["BFsrep09928"],"URL":"https:\/\/doi.org\/10.1038\/srep09928","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,5,21]]},"assertion":[{"value":"17 November 2014","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"23 March 2015","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"21 May 2015","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing financial interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"9928"}}