Simultaneous detection of assembly and disassembly of multivalent HA tag and anti-HA antibody in single in-capillary assay
Herein, we have developed an in‐capillary assay for simultaneous detection of the assembly and disassembly of the multivalent HA tag peptide and antibody. HA tag with hexahistidine at C terminus (YPYDVPDYAG4H6, termed YPYDH6) was conjugated with quantum dots (QDs) by metal‐affinity force to form a m...
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| Veröffentlicht in: | Electrophoresis 2016-08, Vol.37 (15-16), p.2163-2169 |
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| creator | Wang, Jianhao Qin, Yuqin Qin, Haifang Liu, Li Ding, Shumin Teng, Yiwan Ji, Junling Qiu, Lin Jiang, Pengju |
| description | Herein, we have developed an in‐capillary assay for simultaneous detection of the assembly and disassembly of the multivalent HA tag peptide and antibody. HA tag with hexahistidine at C terminus (YPYDVPDYAG4H6, termed YPYDH6) was conjugated with quantum dots (QDs) by metal‐affinity force to form a multivalent HA tag (QD‐YPYDH6). QD‐YPYDH6 and monoclonal anti‐HA antibody (anti‐HA) were sequentially injected into the capillary. They were mixed and assembled inside the capillary. The reaction products were online discriminated and detected by fluorescence coupled capillary electrophoresis (CE‐FL). For the in‐capillary assay, the binding efficiency of the multivalent HA tag and antibody on was influenced by the molar ratio and injection time. Such novel assay could even give out the self‐assembly kinetic constant of QDs and YPYDH6 as KD of 34.1 μM with n (binding cooperativeness) of 2.2 by Hill equation. More importantly, the simultaneous detection of the assembly and imidazole (Im) induced disassembly of the QD‐YPYDH6‐anti‐HA complex was achieved in a single in‐capillary assay. Our study demonstrated a new method for the online detection of antigen‐antibody interactions. |
| doi_str_mv | 10.1002/elps.201600004 |
| format | Article |
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HA tag with hexahistidine at C terminus (YPYDVPDYAG4H6, termed YPYDH6) was conjugated with quantum dots (QDs) by metal‐affinity force to form a multivalent HA tag (QD‐YPYDH6). QD‐YPYDH6 and monoclonal anti‐HA antibody (anti‐HA) were sequentially injected into the capillary. They were mixed and assembled inside the capillary. The reaction products were online discriminated and detected by fluorescence coupled capillary electrophoresis (CE‐FL). For the in‐capillary assay, the binding efficiency of the multivalent HA tag and antibody on was influenced by the molar ratio and injection time. Such novel assay could even give out the self‐assembly kinetic constant of QDs and YPYDH6 as KD of 34.1 μM with n (binding cooperativeness) of 2.2 by Hill equation. More importantly, the simultaneous detection of the assembly and imidazole (Im) induced disassembly of the QD‐YPYDH6‐anti‐HA complex was achieved in a single in‐capillary assay. Our study demonstrated a new method for the online detection of antigen‐antibody interactions.</description><identifier>ISSN: 0173-0835</identifier><identifier>EISSN: 1522-2683</identifier><identifier>DOI: 10.1002/elps.201600004</identifier><identifier>PMID: 27066909</identifier><language>eng</language><publisher>Germany: Blackwell Publishing Ltd</publisher><subject>Anti-HA ; Antibodies ; Antibodies, Monoclonal ; Antigen-Antibody Complex - analysis ; Antigen-antibody interaction ; Assaying ; Assembly ; CE-FL ; Disassembly ; Dismantling ; Electrophoresis, Capillary - methods ; Fluorescence ; Histidine - immunology ; Mathematical analysis ; Oligopeptides - immunology ; Quantum dots ; Quantum Dots - chemistry ; Self assembly ; YPYDVPDYAG4H6</subject><ispartof>Electrophoresis, 2016-08, Vol.37 (15-16), p.2163-2169</ispartof><rights>2016 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim</rights><rights>2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4819-749014323ec4a3e52fd3e489dc3ce13592e98d18396e8b637dd853dbff3e6a513</citedby><cites>FETCH-LOGICAL-c4819-749014323ec4a3e52fd3e489dc3ce13592e98d18396e8b637dd853dbff3e6a513</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Felps.201600004$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Felps.201600004$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27904,27905,45554,45555</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27066909$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wang, Jianhao</creatorcontrib><creatorcontrib>Qin, Yuqin</creatorcontrib><creatorcontrib>Qin, Haifang</creatorcontrib><creatorcontrib>Liu, Li</creatorcontrib><creatorcontrib>Ding, Shumin</creatorcontrib><creatorcontrib>Teng, Yiwan</creatorcontrib><creatorcontrib>Ji, Junling</creatorcontrib><creatorcontrib>Qiu, Lin</creatorcontrib><creatorcontrib>Jiang, Pengju</creatorcontrib><title>Simultaneous detection of assembly and disassembly of multivalent HA tag and anti-HA antibody in single in-capillary assay</title><title>Electrophoresis</title><addtitle>ELECTROPHORESIS</addtitle><description>Herein, we have developed an in‐capillary assay for simultaneous detection of the assembly and disassembly of the multivalent HA tag peptide and antibody. HA tag with hexahistidine at C terminus (YPYDVPDYAG4H6, termed YPYDH6) was conjugated with quantum dots (QDs) by metal‐affinity force to form a multivalent HA tag (QD‐YPYDH6). QD‐YPYDH6 and monoclonal anti‐HA antibody (anti‐HA) were sequentially injected into the capillary. They were mixed and assembled inside the capillary. The reaction products were online discriminated and detected by fluorescence coupled capillary electrophoresis (CE‐FL). For the in‐capillary assay, the binding efficiency of the multivalent HA tag and antibody on was influenced by the molar ratio and injection time. Such novel assay could even give out the self‐assembly kinetic constant of QDs and YPYDH6 as KD of 34.1 μM with n (binding cooperativeness) of 2.2 by Hill equation. More importantly, the simultaneous detection of the assembly and imidazole (Im) induced disassembly of the QD‐YPYDH6‐anti‐HA complex was achieved in a single in‐capillary assay. Our study demonstrated a new method for the online detection of antigen‐antibody interactions.</description><subject>Anti-HA</subject><subject>Antibodies</subject><subject>Antibodies, Monoclonal</subject><subject>Antigen-Antibody Complex - analysis</subject><subject>Antigen-antibody interaction</subject><subject>Assaying</subject><subject>Assembly</subject><subject>CE-FL</subject><subject>Disassembly</subject><subject>Dismantling</subject><subject>Electrophoresis, Capillary - methods</subject><subject>Fluorescence</subject><subject>Histidine - immunology</subject><subject>Mathematical analysis</subject><subject>Oligopeptides - immunology</subject><subject>Quantum dots</subject><subject>Quantum Dots - chemistry</subject><subject>Self assembly</subject><subject>YPYDVPDYAG4H6</subject><issn>0173-0835</issn><issn>1522-2683</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkc1v1DAUxC1ERZfClSPykUsWf8c-VlXZRVqVSgstN8uJXyqDkyxxljb89Tjddq_gy_OTfzPyaBB6R8mSEsI-QtylJSNUkXzEC7SgkrGCKc1fogWhJS-I5vIUvU7px0wYIV6hU1YSpQwxC_RnG9p9HF0H_T5hDyPUY-g73DfYpQRtFSfsOo99SMc9v82a8NtF6Ea8Pseju3ukXDeGIu_zrHo_4dDhFLq7CPlW1G4XYnTDNFu76Q06aVxM8PZpnqFvny6_XqyLzZfV54vzTVELTU1RCkOo4IxDLRwHyRrPQWjja14D5dIwMNpTzY0CXSleeq8l91XTcFBOUn6GPhx8d0P_aw9ptG1INeSfPIa2WSplSZX5H5SSzDKhM7o8oPXQpzRAY3dDaHM4S4mdq7FzNfZYTRa8f_LeVy34I_7cRQbEAbgPEaZ_2NnLzfVW5tBZVhxkIY3wcJS54adVJS-lvb1a2eub7er71e3a3vC_uZ2qTw</recordid><startdate>201608</startdate><enddate>201608</enddate><creator>Wang, Jianhao</creator><creator>Qin, Yuqin</creator><creator>Qin, Haifang</creator><creator>Liu, Li</creator><creator>Ding, Shumin</creator><creator>Teng, Yiwan</creator><creator>Ji, Junling</creator><creator>Qiu, Lin</creator><creator>Jiang, Pengju</creator><general>Blackwell Publishing Ltd</general><scope>BSCLL</scope><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>7X8</scope><scope>7U5</scope><scope>8FD</scope><scope>L7M</scope></search><sort><creationdate>201608</creationdate><title>Simultaneous detection of assembly and disassembly of multivalent HA tag and anti-HA antibody in single in-capillary assay</title><author>Wang, Jianhao ; Qin, Yuqin ; Qin, Haifang ; Liu, Li ; Ding, Shumin ; Teng, Yiwan ; Ji, Junling ; Qiu, Lin ; Jiang, Pengju</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4819-749014323ec4a3e52fd3e489dc3ce13592e98d18396e8b637dd853dbff3e6a513</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Anti-HA</topic><topic>Antibodies</topic><topic>Antibodies, Monoclonal</topic><topic>Antigen-Antibody Complex - analysis</topic><topic>Antigen-antibody interaction</topic><topic>Assaying</topic><topic>Assembly</topic><topic>CE-FL</topic><topic>Disassembly</topic><topic>Dismantling</topic><topic>Electrophoresis, Capillary - methods</topic><topic>Fluorescence</topic><topic>Histidine - immunology</topic><topic>Mathematical analysis</topic><topic>Oligopeptides - immunology</topic><topic>Quantum dots</topic><topic>Quantum Dots - chemistry</topic><topic>Self assembly</topic><topic>YPYDVPDYAG4H6</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Jianhao</creatorcontrib><creatorcontrib>Qin, Yuqin</creatorcontrib><creatorcontrib>Qin, Haifang</creatorcontrib><creatorcontrib>Liu, Li</creatorcontrib><creatorcontrib>Ding, Shumin</creatorcontrib><creatorcontrib>Teng, Yiwan</creatorcontrib><creatorcontrib>Ji, Junling</creatorcontrib><creatorcontrib>Qiu, Lin</creatorcontrib><creatorcontrib>Jiang, Pengju</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Technology Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><jtitle>Electrophoresis</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Jianhao</au><au>Qin, Yuqin</au><au>Qin, Haifang</au><au>Liu, Li</au><au>Ding, Shumin</au><au>Teng, Yiwan</au><au>Ji, Junling</au><au>Qiu, Lin</au><au>Jiang, Pengju</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Simultaneous detection of assembly and disassembly of multivalent HA tag and anti-HA antibody in single in-capillary assay</atitle><jtitle>Electrophoresis</jtitle><addtitle>ELECTROPHORESIS</addtitle><date>2016-08</date><risdate>2016</risdate><volume>37</volume><issue>15-16</issue><spage>2163</spage><epage>2169</epage><pages>2163-2169</pages><issn>0173-0835</issn><eissn>1522-2683</eissn><abstract>Herein, we have developed an in‐capillary assay for simultaneous detection of the assembly and disassembly of the multivalent HA tag peptide and antibody. HA tag with hexahistidine at C terminus (YPYDVPDYAG4H6, termed YPYDH6) was conjugated with quantum dots (QDs) by metal‐affinity force to form a multivalent HA tag (QD‐YPYDH6). QD‐YPYDH6 and monoclonal anti‐HA antibody (anti‐HA) were sequentially injected into the capillary. They were mixed and assembled inside the capillary. The reaction products were online discriminated and detected by fluorescence coupled capillary electrophoresis (CE‐FL). For the in‐capillary assay, the binding efficiency of the multivalent HA tag and antibody on was influenced by the molar ratio and injection time. Such novel assay could even give out the self‐assembly kinetic constant of QDs and YPYDH6 as KD of 34.1 μM with n (binding cooperativeness) of 2.2 by Hill equation. More importantly, the simultaneous detection of the assembly and imidazole (Im) induced disassembly of the QD‐YPYDH6‐anti‐HA complex was achieved in a single in‐capillary assay. Our study demonstrated a new method for the online detection of antigen‐antibody interactions.</abstract><cop>Germany</cop><pub>Blackwell Publishing Ltd</pub><pmid>27066909</pmid><doi>10.1002/elps.201600004</doi><tpages>7</tpages></addata></record> |
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| subjects | Anti-HA Antibodies Antibodies, Monoclonal Antigen-Antibody Complex - analysis Antigen-antibody interaction Assaying Assembly CE-FL Disassembly Dismantling Electrophoresis, Capillary - methods Fluorescence Histidine - immunology Mathematical analysis Oligopeptides - immunology Quantum dots Quantum Dots - chemistry Self assembly YPYDVPDYAG4H6 |
| title | Simultaneous detection of assembly and disassembly of multivalent HA tag and anti-HA antibody in single in-capillary assay |
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