Automated On-tip Affinity Capture Coupled with Mass Spectrometry to Characterize Intact Antibody-Drug Conjugates from Blood
Antibody-drug conjugates (ADCs) present unique challenges for ligand-binding assays primarily due to the dynamic changes of the drug-to-antibody ratio (DAR) distribution in vivo and in vitro. Here, an automated on-tip affinity capture platform with subsequent mass spectrometry analysis was developed...
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| Veröffentlicht in: | Journal of the American Society for Mass Spectrometry 2018-07, Vol.29 (7), p.1532-1537 |
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| container_title | Journal of the American Society for Mass Spectrometry |
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| creator | Li, Ke Sherry Chu, Phillip Y. Fourie-O’Donohue, Aimee Srikumar, Neha Kozak, Katherine R. Liu, Yichin Tran, John C. |
| description | Antibody-drug conjugates (ADCs) present unique challenges for ligand-binding assays primarily due to the dynamic changes of the drug-to-antibody ratio (DAR) distribution in vivo and in vitro. Here, an automated on-tip affinity capture platform with subsequent mass spectrometry analysis was developed to accurately characterize the DAR distribution of ADCs from biological matrices. A variety of elution buffers were tested to offer optimal recovery, with trastuzumab serving as a surrogate to the ADCs. High assay repeatability (CV 3%) was achieved for trastuzumab antibody when captured below the maximal binding capacity of 7.5 μg. Efficient on-tip deglycosylation was also demonstrated in 1 h followed by affinity capture. Moreover, this tip-based platform affords higher throughput for DAR characterization when compared with a well-characterized bead-based method.
Graphical Abstract
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| doi_str_mv | 10.1007/s13361-018-1961-7 |
| format | Article |
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Graphical Abstract
ᅟ</description><identifier>ISSN: 1044-0305</identifier><identifier>EISSN: 1879-1123</identifier><identifier>DOI: 10.1007/s13361-018-1961-7</identifier><identifier>PMID: 29845559</identifier><language>eng</language><publisher>New York: Springer US</publisher><subject>Affinity ; Analytical Chemistry ; Animals ; Antibodies, Monoclonal - blood ; Automation ; Binding ; Bioinformatics ; Biotechnology ; Chemistry ; Chemistry and Materials Science ; Chromatography, Liquid - methods ; Conjugates ; Elution ; Haplorhini ; Humans ; Immunoconjugates - blood ; Immunoconjugates - chemistry ; Mass spectrometry ; Mass Spectrometry - methods ; Monoclonal antibodies ; Organic Chemistry ; Proteomics ; Rats ; Research Article ; Scientific imaging ; Spectroscopy</subject><ispartof>Journal of the American Society for Mass Spectrometry, 2018-07, Vol.29 (7), p.1532-1537</ispartof><rights>American Society for Mass Spectrometry 2018</rights><rights>Journal of The American Society for Mass Spectrometry is a copyright of Springer, (2018). All Rights Reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c372t-d8fc310ad994d0f85785811e55e642948174ec6f68cef76c0a607864758932fc3</citedby><cites>FETCH-LOGICAL-c372t-d8fc310ad994d0f85785811e55e642948174ec6f68cef76c0a607864758932fc3</cites><orcidid>0000-0003-1697-9404</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s13361-018-1961-7$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s13361-018-1961-7$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29845559$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Li, Ke Sherry</creatorcontrib><creatorcontrib>Chu, Phillip Y.</creatorcontrib><creatorcontrib>Fourie-O’Donohue, Aimee</creatorcontrib><creatorcontrib>Srikumar, Neha</creatorcontrib><creatorcontrib>Kozak, Katherine R.</creatorcontrib><creatorcontrib>Liu, Yichin</creatorcontrib><creatorcontrib>Tran, John C.</creatorcontrib><title>Automated On-tip Affinity Capture Coupled with Mass Spectrometry to Characterize Intact Antibody-Drug Conjugates from Blood</title><title>Journal of the American Society for Mass Spectrometry</title><addtitle>J. Am. Soc. Mass Spectrom</addtitle><addtitle>J Am Soc Mass Spectrom</addtitle><description>Antibody-drug conjugates (ADCs) present unique challenges for ligand-binding assays primarily due to the dynamic changes of the drug-to-antibody ratio (DAR) distribution in vivo and in vitro. Here, an automated on-tip affinity capture platform with subsequent mass spectrometry analysis was developed to accurately characterize the DAR distribution of ADCs from biological matrices. A variety of elution buffers were tested to offer optimal recovery, with trastuzumab serving as a surrogate to the ADCs. High assay repeatability (CV 3%) was achieved for trastuzumab antibody when captured below the maximal binding capacity of 7.5 μg. Efficient on-tip deglycosylation was also demonstrated in 1 h followed by affinity capture. Moreover, this tip-based platform affords higher throughput for DAR characterization when compared with a well-characterized bead-based method.
Graphical Abstract
ᅟ</description><subject>Affinity</subject><subject>Analytical Chemistry</subject><subject>Animals</subject><subject>Antibodies, Monoclonal - blood</subject><subject>Automation</subject><subject>Binding</subject><subject>Bioinformatics</subject><subject>Biotechnology</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Chromatography, Liquid - methods</subject><subject>Conjugates</subject><subject>Elution</subject><subject>Haplorhini</subject><subject>Humans</subject><subject>Immunoconjugates - blood</subject><subject>Immunoconjugates - chemistry</subject><subject>Mass spectrometry</subject><subject>Mass Spectrometry - methods</subject><subject>Monoclonal antibodies</subject><subject>Organic Chemistry</subject><subject>Proteomics</subject><subject>Rats</subject><subject>Research Article</subject><subject>Scientific imaging</subject><subject>Spectroscopy</subject><issn>1044-0305</issn><issn>1879-1123</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNp1kc1rFTEUxYNYbK3-AW4k4MZNbJLJ5_I59aNQ6UJdhzSTvM5jZjLmA3n6z5vHqwqFru6B-zvnXjgAvCL4HcFYXmTSdYIgTBQiugn5BJwRJTUihHZPm8aMIdxhfgqe57zDmEis5TNwSrVinHN9Bn5vaomzLX6ANwsq4wo3IYzLWPawt2upycM-1nVq-59juYNfbM7w6-pdSXH2Je1hibC_s8m64tP4y8OrpTQNN0sZb-OwR5epblvGsqvbdibD0Izw_RTj8AKcBDtl__J-noPvHz986z-j65tPV_3mGrlO0oIGFVxHsB20ZgMOikvFFSGecy8Y1UwRybwTQSjngxQOW4GlEkxypTvavOfg7TF3TfFH9bmYeczOT5NdfKzZUMwk5YQr2tA3D9BdrGlp3x0o0QlBCW8UOVIuxZyTD2ZN42zT3hBsDs2YYzOmNWMOzRjZPK_vk-vt7Id_jr9VNIAegdxWy9an_6cfT_0DidKYsg</recordid><startdate>20180701</startdate><enddate>20180701</enddate><creator>Li, Ke Sherry</creator><creator>Chu, Phillip Y.</creator><creator>Fourie-O’Donohue, Aimee</creator><creator>Srikumar, Neha</creator><creator>Kozak, Katherine R.</creator><creator>Liu, Yichin</creator><creator>Tran, John C.</creator><general>Springer US</general><general>Springer Nature B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FE</scope><scope>8FG</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>M2O</scope><scope>MBDVC</scope><scope>P5Z</scope><scope>P62</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0003-1697-9404</orcidid></search><sort><creationdate>20180701</creationdate><title>Automated On-tip Affinity Capture Coupled with Mass Spectrometry to Characterize Intact Antibody-Drug Conjugates from Blood</title><author>Li, Ke Sherry ; Chu, Phillip Y. ; Fourie-O’Donohue, Aimee ; Srikumar, Neha ; Kozak, Katherine R. ; Liu, Yichin ; Tran, John C.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c372t-d8fc310ad994d0f85785811e55e642948174ec6f68cef76c0a607864758932fc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Affinity</topic><topic>Analytical Chemistry</topic><topic>Animals</topic><topic>Antibodies, Monoclonal - blood</topic><topic>Automation</topic><topic>Binding</topic><topic>Bioinformatics</topic><topic>Biotechnology</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>Chromatography, Liquid - methods</topic><topic>Conjugates</topic><topic>Elution</topic><topic>Haplorhini</topic><topic>Humans</topic><topic>Immunoconjugates - blood</topic><topic>Immunoconjugates - chemistry</topic><topic>Mass spectrometry</topic><topic>Mass Spectrometry - methods</topic><topic>Monoclonal antibodies</topic><topic>Organic Chemistry</topic><topic>Proteomics</topic><topic>Rats</topic><topic>Research Article</topic><topic>Scientific imaging</topic><topic>Spectroscopy</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Li, Ke Sherry</creatorcontrib><creatorcontrib>Chu, Phillip Y.</creatorcontrib><creatorcontrib>Fourie-O’Donohue, Aimee</creatorcontrib><creatorcontrib>Srikumar, Neha</creatorcontrib><creatorcontrib>Kozak, Katherine R.</creatorcontrib><creatorcontrib>Liu, Yichin</creatorcontrib><creatorcontrib>Tran, John C.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Research Library (Alumni Edition)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>ProQuest Central Essentials</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>Research Library Prep</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Research Library</collection><collection>Research Library (Corporate)</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of the American Society for Mass Spectrometry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Li, Ke Sherry</au><au>Chu, Phillip Y.</au><au>Fourie-O’Donohue, Aimee</au><au>Srikumar, Neha</au><au>Kozak, Katherine R.</au><au>Liu, Yichin</au><au>Tran, John C.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Automated On-tip Affinity Capture Coupled with Mass Spectrometry to Characterize Intact Antibody-Drug Conjugates from Blood</atitle><jtitle>Journal of the American Society for Mass Spectrometry</jtitle><stitle>J. Am. Soc. Mass Spectrom</stitle><addtitle>J Am Soc Mass Spectrom</addtitle><date>2018-07-01</date><risdate>2018</risdate><volume>29</volume><issue>7</issue><spage>1532</spage><epage>1537</epage><pages>1532-1537</pages><issn>1044-0305</issn><eissn>1879-1123</eissn><abstract>Antibody-drug conjugates (ADCs) present unique challenges for ligand-binding assays primarily due to the dynamic changes of the drug-to-antibody ratio (DAR) distribution in vivo and in vitro. Here, an automated on-tip affinity capture platform with subsequent mass spectrometry analysis was developed to accurately characterize the DAR distribution of ADCs from biological matrices. A variety of elution buffers were tested to offer optimal recovery, with trastuzumab serving as a surrogate to the ADCs. High assay repeatability (CV 3%) was achieved for trastuzumab antibody when captured below the maximal binding capacity of 7.5 μg. Efficient on-tip deglycosylation was also demonstrated in 1 h followed by affinity capture. Moreover, this tip-based platform affords higher throughput for DAR characterization when compared with a well-characterized bead-based method.
Graphical Abstract
ᅟ</abstract><cop>New York</cop><pub>Springer US</pub><pmid>29845559</pmid><doi>10.1007/s13361-018-1961-7</doi><tpages>6</tpages><orcidid>https://orcid.org/0000-0003-1697-9404</orcidid></addata></record> |
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| subjects | Affinity Analytical Chemistry Animals Antibodies, Monoclonal - blood Automation Binding Bioinformatics Biotechnology Chemistry Chemistry and Materials Science Chromatography, Liquid - methods Conjugates Elution Haplorhini Humans Immunoconjugates - blood Immunoconjugates - chemistry Mass spectrometry Mass Spectrometry - methods Monoclonal antibodies Organic Chemistry Proteomics Rats Research Article Scientific imaging Spectroscopy |
| title | Automated On-tip Affinity Capture Coupled with Mass Spectrometry to Characterize Intact Antibody-Drug Conjugates from Blood |
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