Humanized Bispecific Antibody (mPEG×HER2) Rapidly Confers PEGylated nanoaparticles Tumor Specificity for Multimodality Imaging in Breast Cancer

Humanized Bispecific Antibody (mPEG×HER2) Rapidly Confers PEGylated nanoaparticles Tumor Specificity for Multimodality Imaging in Breast Cancer | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Help Center Sign In Submit a Preprint Cite Share Download PDF Research Humanized Bispecific Antibody (mPEG×HER2) Rapidly Confers PEGylated nanoaparticles Tumor Specificity for Multimodality Imaging in Breast Cancer Yi-An Cheng, Tung-Ho Wu, Yun-Ming Wang, Tian-Lu Cheng, I-Ju Chen, and 9 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-28604/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 27 Aug, 2020 Read the published version in Journal of Nanobiotechnology → Version 1 posted 8 You are reading this latest preprint version Abstract Background: Developing a universal strategy to improve the specificity and sensitivity of PEGylated nanoaparticles (PEG-NPs) for assisting in the diagnosis of tumors is important in multimodality imaging. Here, we developed the anti-methoxypolyethylene glycol (mPEG) bispecific antibody (BsAb; mPEG×HER2), which has dual specificity for mPEG and human epidermal growth factor receptor 2 (HER2), with a diverse array of PEG-NPs to confer nanoparticles with HER2 specificity and stronger intensity. Result: We used a one-step formulation to rapidly modify the nanoprobes with mPEG×HER2 and optimized the modified ratio of BsAbs on several PEG-NPs (Lipo-DiR, SPIO, Qdot and AuNP). The αHER2/PEG-NPs could specifically target MCF7/HER2 cells (HER2+) but not MCF7/neo1 cells (HER2-). The αHER2/Lipo-DiR and αHER2/SPIO could enhance the sensitivity of untargeted PEG-NPs on MCF7/HER2 (HER2+). In in vivo imaging, αHER2/Lipo-DiR and αHER2/SPIO increased the specific targeting and enhanced mPEG-nanoprobe accumulation 161% and 187%, respectively, in HER2-overexpressing tumors. Conclusion: mPEG×HER2, therefore, provided a simple one-step formulation to confer HER2-specific targeting and enhanced sensitivity and contrast intensity on HER2 positive tumors for multimodality imaging. Nanoscience Bispecific Antibody PEGylated nanocparticle nano-contrast agent multimodality image polyethylene glycol anti-PEG antibody one-step formulation Tumor Specificity targeted image Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Full Text Supplementary Files ST11.tif Cite Share Download PDF Status: Published Journal Publication published 27 Aug, 2020 Read the published version in Journal of Nanobiotechnology → Version 1 posted Reviewer # 2 agreed at journal 13 Jun, 2020 Review # 1 received at journal 07 Jun, 2020 Reviewer # 1 agreed at journal 25 May, 2020 Reviewers invited by journal 23 May, 2020 Editor assigned by journal 12 May, 2020 First submitted to journal 11 May, 2020 Submission checks completed at journal 11 May, 2020 Editor invited by journal 11 May, 2020 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-28604","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research","associatedPublications":[],"authors":[{"id":579763,"identity":"413924c3-3641-4e81-a53c-5bc49fd64c4a","order_by":1,"name":"Yi-An Cheng","email":"","orcid":"","institution":"Kaohsiung Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yi-An","middleName":"","lastName":"Cheng","suffix":""},{"id":579764,"identity":"75bc0a0a-f88b-4036-b9e7-b9f2c1ac19ed","order_by":2,"name":"Tung-Ho Wu","email":"","orcid":"","institution":"Kaohsiung Veterans General Hospital","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Tung-Ho","middleName":"","lastName":"Wu","suffix":""},{"id":579765,"identity":"2c22aa16-2d91-471d-a949-4957859ff1b1","order_by":3,"name":"Yun-Ming Wang","email":"","orcid":"","institution":"National Chiao Tung University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yun-Ming","middleName":"","lastName":"Wang","suffix":""},{"id":579766,"identity":"43ee8f3c-ac7b-4180-a614-989c72c6ced0","order_by":4,"name":"Tian-Lu Cheng","email":"","orcid":"https://orcid.org/0000-0002-7263-2766","institution":"Kaohsiung Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Tian-Lu","middleName":"","lastName":"Cheng","suffix":""},{"id":579767,"identity":"96f6c40a-e88d-40fc-bb51-4bb1c77699b9","order_by":5,"name":"I-Ju Chen","email":"","orcid":"","institution":"Kaohsiung Medical 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Chen","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAs0lEQVRIiWNgGAWjYBAC9gYeIFmRAOZIEKWF5wBIyxm4FgMitTC2kaSF/ezBx4Xz0qINDjAfvM3D8CexgaAWnrxk45nbcnI3HGBLtuZhMCCsxV6Cx0yad1sFUAuQAdSSS9gWCR7z37xzQFr4vxGtxYyZtwHkMB42IrXw5BhL8xxLy515mM3Yco6BcT1hLexnDD/z1CTn9h1vfnjjTYWcMSEdSIAZRBARLaNgFIyCUTAKiAAAc7A0v3M/6B8AAAAASUVORK5CYII=","orcid":"","institution":"","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Fang-Ming","middleName":"","lastName":"Chen","suffix":""}],"badges":[],"createdAt":"2020-05-12 12:02:02","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-28604/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-28604/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1186/s12951-020-00680-9","type":"published","date":"2020-08-27T12:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":1116084,"identity":"81529b0a-9f16-45c6-b7db-1a50a8db3886","added_by":"auto","created_at":"2020-05-17 20:40:23","extension":"tif","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":552338,"visible":true,"origin":"","legend":"Humanized Bispecific Antibody (mPEG×HER2) Rapidly Confers PEG-NPs Tumor Specificity for Multimodality Imaging in Breast Cancer\nanti-mPEG BsAbs (mPEG×markers) provide an easy, universal and one-step formulation for any naive PEG-NPs to accelerate the development of targeted PEG-NPs for multimodality imaging in the clinic.","description":"","filename":"1.tif","url":"https://assets-eu.researchsquare.com/files/rs-28604/v1/1.tif"},{"id":1116086,"identity":"7fcc63e1-433f-4f87-ad8f-8aa8b4168ebc","added_by":"auto","created_at":"2020-05-17 20:40:23","extension":"tif","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":192344,"visible":true,"origin":"","legend":"The HER2 binding ability of αHER2/PEG-NPs\n \nDifferent BsAb:mPEG ratios of (A) αHER2/Lipo-DiR, (B) αHER2/SPIO, (C) αHER2/Qdot and (D) αHER2/AuNP were incubated with MCF7/HER2, and then anti-PEG antibody was added to detect PEG-NPs via ELISA (n=3, triplicate). Bars, SD.","description":"","filename":"21.tif","url":"https://assets-eu.researchsquare.com/files/rs-28604/v1/21.tif"},{"id":1116087,"identity":"f8c7224b-2120-42b0-96ad-2df3621ee82a","added_by":"auto","created_at":"2020-05-17 20:40:23","extension":"tif","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":179447,"visible":true,"origin":"","legend":"Specificity of αHER2/PEG-NPs for HER2+ cancer cells\n \nMCF7/HER2 (HER+) (circle shape) and MCF7/neo1(HER2-) (square shape) cancer cells in 96-well plates were incubated with mPEG×HER2 (solid shape) and mPEG×DNS (hollow shape) modified with different contrast agents. After washing, bound contrast agents were detected by ELISA (n=3, triplicate). Bars, SD.","description":"","filename":"31.tif","url":"https://assets-eu.researchsquare.com/files/rs-28604/v1/31.tif"},{"id":1116088,"identity":"cb940489-5a6d-4400-8268-c90c44ba3893","added_by":"auto","created_at":"2020-05-17 20:40:23","extension":"tif","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":420320,"visible":true,"origin":"","legend":"In vitro sensitivity image of αHER2/PEG-NPs\nMCF7/HER2 (HER+) cancer cells incubated with HER2 targeted-contrast agent serial dilution concentration. (A) αHER2/Lipo-DiR, αDNS/Lipo-DiR and Lipo-DiR was added to cells. Fluorescence images were obtained by the IVIS spectrum system. (B) The result was the calculation of average radiant efficiency of (A). (C) αHER2/SPIO, αDNS/SPIO and SPIO was added to cells. MR imaging was performed with a 7.0 T MR imaging scanner. (D) The result from (C) was calculated by [treated SI-untreated SI]/ untreated SI*100. *P \u003c 0.05, **P \u003c 0.01, ***P \u003c 0.001, ****P \u003c 0.0001(unpaired t test).","description":"","filename":"42.tif","url":"https://assets-eu.researchsquare.com/files/rs-28604/v1/42.tif"},{"id":1116089,"identity":"91a583ee-b5e1-40d6-8476-651d10dc2d61","added_by":"auto","created_at":"2020-05-17 20:40:23","extension":"tif","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":813928,"visible":true,"origin":"","legend":"In vivo IVIS imaging of αHER2/SPIO and αDNS/SPIO\nThe tumor delivery of αHER2/PLD in HER2-overexpressing tumors. (A) αHER2/Lipo-DiR and Lipo- DiR were intravenously injected in mice bearing HER2high (left flank) and HER2low (right flank) tumors. The fluorescence intensity of DiR was detected at 24 h and 72 h after injection by IVIS. (B) Quantification of average radiant efficiency in HER2+ tumor and HER2- tumor at 72 h.","description":"","filename":"51.tif","url":"https://assets-eu.researchsquare.com/files/rs-28604/v1/51.tif"},{"id":1116090,"identity":"a9f97d31-04b2-4306-b351-cdda813eb8fb","added_by":"auto","created_at":"2020-05-17 20:40:24","extension":"tif","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":460499,"visible":true,"origin":"","legend":"In vivo MR imaging of αHER2/SPIO and αDNS/SPIO\nNude mice bearing MCF7/HER2 (right m.f.p) and MCF7/neo1 (left m.f.p) tumors were intravenously injected with αHER2/SPIO and αDNS/ SPIO (10 mg/kg). (A) Mice were sequentially imaged at pre- treatment and 24 h with a 7.0 T MR imaging scanner. (B) The result was calculated as the percentage of negative signal enhancement by [pretreated signal intensity (SI0)-treated signal intensity (SI24)]/SI0*100.","description":"","filename":"6.tif","url":"https://assets-eu.researchsquare.com/files/rs-28604/v1/6.tif"},{"id":1116091,"identity":"378f6892-aaa8-46ed-b5be-6af1e4432f6d","added_by":"auto","created_at":"2020-05-17 20:40:24","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1555151,"visible":true,"origin":"","legend":"","description":"","filename":"SubmtionBMCnanobiotechnologyimagepaper.pdf","url":"https://assets-eu.researchsquare.com/files/rs-28604/v1_stamped.pdf"},{"id":1116085,"identity":"0dadea51-c683-4137-b4f2-9dc7548d64ee","added_by":"auto","created_at":"2020-05-17 20:40:23","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1474809,"visible":true,"origin":"","legend":"","description":"","filename":"SubmtionBMCnanobiotechnologyimagepaper.pdf","url":"https://assets-eu.researchsquare.com/files/rs-28604/v1/SubmtionBMCnanobiotechnologyimagepaper.pdf"},{"id":13504016,"identity":"16c9664a-61dc-47d3-9fe5-b20eca9d88d1","added_by":"auto","created_at":"2021-09-16 23:21:29","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2331742,"visible":true,"origin":"","legend":"","description":"","filename":"SubmtionBMCnanobiotechnologyimagepaper.pdf","url":"https://assets-eu.researchsquare.com/files/rs-28604/v1_covered.pdf"},{"id":1116083,"identity":"445ed0be-aa4b-4141-a413-1dbfe4d44bea","added_by":"auto","created_at":"2020-05-17 20:40:23","extension":"tif","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":237392,"visible":true,"origin":"","legend":"","description":"","filename":"ST11.tif","url":"https://assets-eu.researchsquare.com/files/rs-28604/v1/ST11.tif"}],"financialInterests":"","formattedTitle":"Humanized Bispecific Antibody (mPEG×HER2) Rapidly Confers PEGylated nanoaparticles Tumor Specificity for Multimodality Imaging in Breast Cancer","fulltext":[{"header":"Full Text","content":"\u003cp\u003eThis preprint is available for \u003ca href='/article/rs-28604/latest.pdf' target='_blank'\u003edownload as a PDF\u003c/a\u003e.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"journal-of-nanobiotechnology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jnan","sideBox":"Learn more about [Journal of Nanobiotechnology](http://jnanobiotechnology.biomedcentral.com)","snPcode":"12951","submissionUrl":"https://submission.nature.com/new-submission/12951/3","title":"Journal of Nanobiotechnology","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Bispecific Antibody, PEGylated nanocparticle, nano-contrast agent, multimodality image, polyethylene glycol, anti-PEG antibody, one-step formulation, Tumor Specificity, targeted image","lastPublishedDoi":"10.21203/rs.3.rs-28604/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-28604/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBackground: Developing a universal strategy to improve the specificity and sensitivity of PEGylated nanoaparticles (PEG-NPs) for assisting in the diagnosis of tumors is important in multimodality imaging. Here, we developed the anti-methoxypolyethylene glycol (mPEG) bispecific antibody (BsAb; mPEG×HER2), which has dual specificity for mPEG and human epidermal growth factor receptor 2 (HER2), with a diverse array of PEG-NPs to confer nanoparticles with HER2 specificity and stronger intensity. \u003c/p\u003e\u003cp\u003eResult: We used a one-step formulation to rapidly modify the nanoprobes with mPEG×HER2 and optimized the modified ratio of BsAbs on several PEG-NPs (Lipo-DiR, SPIO, Qdot and AuNP). The αHER2/PEG-NPs could specifically target MCF7/HER2 cells (HER2+) but not MCF7/neo1 cells (HER2-). The αHER2/Lipo-DiR and αHER2/SPIO could enhance the sensitivity of untargeted PEG-NPs on MCF7/HER2 (HER2+). In in vivo imaging, αHER2/Lipo-DiR and αHER2/SPIO increased the specific targeting and enhanced mPEG-nanoprobe accumulation 161% and 187%, respectively, in HER2-overexpressing tumors. \u003c/p\u003e\u003cp\u003eConclusion: mPEG×HER2, therefore, provided a simple one-step formulation to confer HER2-specific targeting and enhanced sensitivity and contrast intensity on HER2 positive tumors for multimodality imaging.\u003c/p\u003e","manuscriptTitle":"Humanized Bispecific Antibody (mPEG×HER2) Rapidly Confers PEGylated nanoaparticles Tumor Specificity for Multimodality Imaging in Breast Cancer","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2020-05-17 20:40:22","doi":"10.21203/rs.3.rs-28604/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2020-06-13T12:00:00+00:00","index":2,"fulltext":""},{"type":"editorInvitedReview","content":"","date":"2020-06-07T12:00:00+00:00","index":1,"fulltext":"Recommendation: Reviewer's comments unavailable due to the journal's policy.\n"},{"type":"reviewerAgreed","content":"","date":"2020-05-25T12:00:00+00:00","index":1,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2020-05-23T12:00:00+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2020-05-12T12:00:00+00:00","index":"","fulltext":""},{"type":"submitted","content":"","date":"2020-05-11T12:00:00+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2020-05-11T12:00:00+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2020-05-11T12:00:00+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-nanobiotechnology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jnan","sideBox":"Learn more about [Journal of Nanobiotechnology](http://jnanobiotechnology.biomedcentral.com)","snPcode":"12951","submissionUrl":"https://submission.nature.com/new-submission/12951/3","title":"Journal of Nanobiotechnology","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"581fcfa9-89d6-484d-bbbd-2b3e8b1311e4","owner":[],"postedDate":"May 17th, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":101578,"name":"Nanoscience"}],"tags":[],"updatedAt":"2020-09-03T15:58:30+00:00","versionOfRecord":{"articleIdentity":"rs-28604","link":"https://doi.org/10.1186/s12951-020-00680-9","journal":{"identity":"journal-of-nanobiotechnology","isVorOnly":false,"title":"Journal of Nanobiotechnology"},"publishedOn":"2020-08-27 12:00:00","publishedOnDateReadable":"August 27th, 2020"},"versionCreatedAt":"2020-05-17 20:40:22","video":"","vorDoi":"10.1186/s12951-020-00680-9","vorDoiUrl":"https://doi.org/10.1186/s12951-020-00680-9","workflowStages":[]},"version":"v1","identity":"rs-28604","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-28604","identity":"rs-28604","version":["v1"]},"buildId":"GqpaHPwrfC8PjnIFayRh5","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}