Multiplexed Continuous Biosensing by Single-Molecule Encoded Nanoswitches

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usa-input-error-message selection-validation-message"> Name must be less than 100 characters </div> </div> <div class="action-panel-control-wrap existing-collections-controls"> <label for="collections-action-panel-add-to-existing" class="action-panel-label"> Choose a collection: </label> <select id="collections-action-panel-add-to-existing" class="action-panel-selector collections-action-add-to-existing" data-ga-category="save_share" data-ga-action="select_collection" data-ga-label="($('#collections-action-add-to-existing').val() === 'Favorites') ? 'Favorites' : 'non_favorites_collection'"> </select> <div class="collections-retry-load-on-error usa-input-error-message selection-validation-message"> Unable to load your collection due to an error<br> <a href="#">Please try again</a> </div> </div> </div> <div class="action-panel-actions"> <button class="action-panel-submit" type="submit" data-loading-label="Adding..." data-pinger-ignore data-ga-category="save_share" data-ga-action="collections" data-ga-label="add"> Add </button> <button class="action-panel-cancel" aria-label="Close 'Add to Collections' panel" ref="linksrc=close_collections_panel" aria-controls="collections-action-panel" aria-expanded="false" data-ga-category="save_share" data-ga-action="collections" data-ga-label="cancel"> Cancel </button> </div> </form> </div> </div> <div id="bibliography-action-panel" class="bibliography-action-panel action-panel in-progress-dots-panel" aria-hidden="true" data-bibliography-open-panel-enabled="false" data-bibliography-open-panel-url-hash="#open-bibliography-panel"> <div class="inner-wrap"> <h3 class="action-panel-heading"> Add to My Bibliography </h3> <form id="bibliography-action-panel-form" class="bibliography-action-panel-form action-panel-content action-form action-panel-smaller-selectors" data-add-to-bibliography-max-amount="100" data-add-to-bibliography-batch-size="10" data-bibliography-delegates-url="/list-bibliography-delegates/" data-add-to-bibliography-url="/add-to-bibliography/" data-get-article-ids-by-search-url="/get-article-ids-by-search/" data-mybib-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/mybibliography/"> <input type="hidden" name="csrfmiddlewaretoken" value="7hJMujUN5Gw1LR8mQES9HbOXgpoyEdRQ0hVCGPA1xVkOOEhUUhs2KhKE3SqXkiBC"> <div class="action-panel-control-wrap bibliographies-controls"> <div class="choice-group"> <ul class="bibliographies-action-add radio-group-items"> <li> <input name="bibliography" id="my-bibliography" class="my-bibliography" type="radio" checked/> <label for="my-bibliography">My Bibliography</label> </li> </ul> </div> </div> <div class="bibliographies-retry-load-on-error usa-input-error-message selection-validation-message"> Unable to load your delegates due to an error<br> <a href="#">Please try again</a> </div> <div class="action-panel-actions"> <button class="action-panel-submit" type="submit" data-loading-label="Adding..." data-pinger-ignore> Add </button> <button class="action-panel-cancel" aria-label="Close 'Add to bibliography' panel" ref="linksrc=close_bibliography_panel" aria-controls="bibliography-action-panel" aria-expanded="false" data-ga-category="save_share" data-ga-action="mybib" data-ga-label="cancel"> Cancel </button> </div> </form> </div> </div> <div id="saved-search-action-panel" class="saved-search-action-panel action-panel " aria-hidden="true" data-saved-search-open-panel-enabled="false" data-saved-search-open-panel-url-hash="#open-saved-search-panel"> <div class="inner-wrap"> <h2 class="action-panel-heading"> Your saved search </h2> <form id="saved-search-action-panel-form" class="saved-search-action-panel-form action-panel-content action-form" data-create-saved-search-url="/create-saved-search/" data-try-search-terms-url="/try-search-term/" data-saved-search-root-url="https://www.ncbi.nlm.nih.gov/myncbi/searches/"> <input type="hidden" name="csrfmiddlewaretoken" value="7hJMujUN5Gw1LR8mQES9HbOXgpoyEdRQ0hVCGPA1xVkOOEhUUhs2KhKE3SqXkiBC"> <div class="action-panel-control-wrap"> <label for="saved-search-name" class="action-panel-label saved-search-name-label required-field-asterisk"> Name of saved search: </label> <input maxlength="200" type="text" name="saved-search-name" id="saved-search-name" class="saved-search-name" value="" required pattern="[^"&=<>\/]*" title="The following characters are not allowed in the Name field: "&=<>/"> </div> <div class="action-panel-control-wrap"> <label for="saved-search-term" class="action-panel-label required-field-asterisk"> Search terms: </label> <textarea name="saved-search-term" id="saved-search-term" class="saved-search-term" required></textarea> </div> <div class="test-search-term-wrap"> <a href="#" class="try-search-term">Test search terms</a> </div> <div class="choice-group action-panel-extra-margin-top"> <span class="action-panel-label" id="fieldset-label"> Would you like email updates of new search results? </span> <fieldset id="saved-search-alert" aria-describedby="fieldset-label"> <legend class="usa-sr-only">Saved Search Alert Radio Buttons</legend> <ul class="radio-group-items"> <li> <input type="radio" id="saved-search-alert-yes" class="saved-search-alert-yes" name="saved-search-alert" value="yes" checked> <label for="saved-search-alert-yes" class="action-panel-label">Yes</label> </li> <li> <input aria-label="No radio input" type="radio" id="saved-search-alert-no" class="saved-search-alert-no" name="saved-search-alert" value="no"> <label for="saved-search-alert-no" class="action-panel-label">No</label> </li> </ul> </fieldset> </div> <div class="alert-schedule-wrap"> <div class="action-panel-control-wrap"> <label class="action-panel-label"> Email: </label> <span aria-label="Email address" id="saved-search-email" class="action-panel-label"><span class="action-panel-label-bold"></span> (<a class="myncbi-account-settings" href="https://www.ncbi.nlm.nih.gov/account/settings/">change</a>)</span> </div> <div class="action-panel-control-wrap action-panel-extra-margin-top"> <label for="saved-search-frequency" class="action-panel-label"> Frequency: </label> <select id="saved-search-frequency" class="no-border-panel-selector saved-search-frequency"> <option value="monthly">Monthly</option> <option value="weekly">Weekly</option> <option value="daily">Daily</option> </select> </div> <div class="action-panel-control-wrap saved-search-monthly-additional"> <label for="saved-search-monthly-on-day" class="action-panel-label"> Which day? </label> <select id="saved-search-monthly-on-day" class="no-border-panel-selector"> <option value="Sunday">The first Sunday</option> <option value="Monday">The first Monday</option> <option value="Tuesday">The first Tuesday</option> <option value="Wednesday">The first Wednesday</option> <option value="Thursday">The first Thursday</option> <option value="Friday">The first Friday</option> <option value="Saturday">The first Saturday</option> <option value="day">The first day</option> <option value="weekday">The first weekday</option> </select> </div> <div class="action-panel-control-wrap saved-search-weekly-additional"> <label for="saved-search-weekly-on-day" class="action-panel-label"> Which day? </label> <select id="saved-search-weekly-on-day" class="no-border-panel-selector saved-search-weekly-on-day"> <option value="Sunday">Sunday</option> <option value="Monday">Monday</option> <option value="Tuesday">Tuesday</option> <option value="Wednesday">Wednesday</option> <option value="Thursday">Thursday</option> <option value="Friday">Friday</option> <option value="Saturday">Saturday</option> </select> </div> <div class="action-panel-control-wrap"> <label for="saved-search-report" class="action-panel-label"> Report format: </label> <select id="saved-search-report" class="no-border-panel-selector saved-search-report"> <option value="DocSum">Summary</option> <option value="DocSumText">Summary (text)</option> <option value="Abstract">Abstract</option> <option value="AbstractText">Abstract (text)</option> <option value="MEDLINE">PubMed</option> </select> </div> <div class="action-panel-control-wrap"> <label for="saved-search-amount" class="action-panel-label"> Send at most: </label> <select id="saved-search-amount" class="no-border-panel-selector saved-search-amount"> <option value="1">1 item</option> <option value="5" selected>5 items</option> <option value="10">10 items</option> <option value="20">20 items</option> <option value="50">50 items</option> <option value="100">100 items</option> <option value="200">200 items</option> </select> </div> <div> <input type="checkbox" id="saved-search-send-if-no-result" class="saved-search-send-if-no-result" name="saved-search-send-if-no-result"> <label for="saved-search-send-if-no-result" class="action-panel-label smaller-checkbox"> Send even when there aren't any new results </label> </div> <div class="action-panel-control-wrap option-text-in-email-wrap"> <label for="saved-search-email-text" class="action-panel-label"> Optional text in email: </label> <textarea name="saved-search-email-text" id="saved-search-email-text" class="saved-search-email-text"></textarea> </div> </div> <div class="action-panel-actions"> <button class="action-panel-submit" type="submit" data-loading-label="Saving..." data-ga-category="save_share" data-ga-action="alert" data-ga-label="save"> Save </button> <button class="action-panel-cancel" aria-label="Close 'Your saved search' panel" ref="linksrc=close_saved_search_panel" aria-controls="saved-search-action-panel" aria-expanded="false" data-ga-category="save_share" data-ga-action="alert" data-ga-label="cancel"> Cancel </button> </div> </form> </div> </div> <div id="citation-manager-action-panel" class="citation-manager-action-panel action-panel" aria-hidden="true"> <div class="inner-wrap"> <h2 class="action-panel-heading"> Create a file for external citation management software </h2> <form id="citation-manager-action-panel-form" class="action-panel-content action-form" action="/results-export-ids/" data-by-search-action="/results-export-search-data/" data-by-ids-action="/results-export-ids/" method="post" data-by-search-method="post" data-by-ids-method="post"> <input type="hidden" name="csrfmiddlewaretoken" value="7hJMujUN5Gw1LR8mQES9HbOXgpoyEdRQ0hVCGPA1xVkOOEhUUhs2KhKE3SqXkiBC"> <input name="results-format" type="hidden" value="pubmed"/> <div class="action-panel-actions"> <button class="action-panel-submit" type="submit" data-loading-label="Sending..." data-ga-category="save_share" data-ga-action="citation_manager" data-ga-label="save"> Create file </button> <button class="action-panel-cancel" aria-label="Close 'Send citations to citation manager' panel" ref="linksrc=close_citation_manager_panel" aria-controls="citation-manager-action-panel" aria-expanded="false" data-ga-category="save_share" data-ga-action="citation_manager" data-ga-label="cancel"> Cancel </button> </div> </form> </div> </div> <div id="rss-action-panel" class="rss-action-panel action-panel " aria-hidden="true"> <div class="inner-wrap"> <h2 class="action-panel-heading"> Your RSS Feed </h2> <form id="rss-action-panel-form" class="rss-action-panel-form action-panel-content action-form" data-create-rss-feed-url="/create-rss-feed-url/" data-search-form-term-value=""> <input type="hidden" name="csrfmiddlewaretoken" value="7hJMujUN5Gw1LR8mQES9HbOXgpoyEdRQ0hVCGPA1xVkOOEhUUhs2KhKE3SqXkiBC"> <div class="action-panel-control-wrap"> <label for="rss-name" class="action-panel-label required-field-asterisk"> Name of RSS Feed: </label> <input maxlength="200" placeholder="Name" type="text" name="rss-name" id="rss-name" class="rss-name" value='' required pattern="[^"&=<>\/]*" title="The following characters are not allowed in the Name field: "&=<>/"> </div> <div class="rss-limit-wrap"> <div class="action-panel-control-wrap action-panel-extra-margin-top"> <label for="rss-limit" class="action-panel-label"> Number of items displayed: </label> <select id="rss-limit" class="no-border-panel-selector rss-limit"> <option value="5">5</option> <option value="10">10</option> <option value="15" selected="selected">15</option> <option value="20">20</option> <option value="50">50</option> <option value="100">100</option> </select> </div> </div> <div class="action-panel-actions"> <button class="action-panel-submit" type="submit" data-loading-label="Creating..." data-ga-category="save_share" data-ga-action="alert" data-ga-label="save"> Create RSS </button> <button class="action-panel-cancel" aria-label="Close 'Your RSS' panel" ref="linksrc=close_rss_panel" aria-controls="rss-action-panel" aria-expanded="false" data-ga-category="save_share" data-ga-action="alert" data-ga-label="cancel"> Cancel </button> </div> <div class="action-panel-control-wrap rss-link-copy-wrap"> <label for="rss-link" class="usa-sr-only">RSS Link</label> <input placeholder="Your RSS Feed Link" type="text" name="rss-link" id="rss-link" class="rss-link" title="RSS Link"> <button type="button" disabled class="rss-link-copy-button disabled" data-ga-category="save_share" data-ga-action="rss" data-ga-label="copy"> Copy </button> </div> </form> </div> </div> </div> </div> <div class="article-page" id="article-page" data-article-pmid="32091908"> <aside class="page-sidebar"> <div class="inner-wrap"> <div class="full-text-links"> <div class="full-view"> <h3 class="title"> Full text links </h3> <div class="full-text-links-list"> <a class="link-item dialog-focus" 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required-field-asterisk"> Name your collection: </label><input type="text" name="add-to-new-collection" id="collections-action-dialog-add-to-new" class="collections-action-add-to-new" pattern="[^"&=<>\/]*" title="The following characters are not allowed in the Name field: "&=<>/" maxlength="100" data-ga-category="collections_button" data-ga-action="create_collection" data-ga-label="non_favorties_collection"><div class="collections-new-name-too-long usa-input-error-message selection-validation-message"> Name must be less than 100 characters </div></div><div class="action-panel-control-wrap existing-collections-controls"><label for="collections-action-dialog-add-to-existing" class="action-panel-label"> Choose a collection: </label><select id="collections-action-dialog-add-to-existing" class="action-panel-selector collections-action-add-to-existing" data-ga-category="collections_button" data-ga-action="select_collection" data-ga-label="($('.collections-action-add-to-existing').val() === 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'Favorites' : 'non_favorites_collection'"></select><div class="collections-retry-load-on-error usa-input-error-message selection-validation-message"> Unable to load your collection due to an error<br><a href="#">Please try again</a></div></div></div><div class="action-panel-actions"><button class="action-panel-submit" type="submit" data-loading-label="Adding..." data-pinger-ignore data-ga-category="collections_button" data-ga-action="click" data-ga-label="add"> Add </button><button class="action-panel-cancel" aria-label="Close 'Add to Collections' panel" ref="linksrc=close_collections_panel" aria-controls="collections-action-panel" aria-expanded="false" data-ga-category="collections_button" data-ga-action="click" data-ga-label="cancel"> Cancel </button></div></form></div></div></div></div><button class="more-actions-button more-actions-dialog-trigger" title="Open dialog with more actions to take" ref="linksrc=more_actions_btn"></button><div class="display-options" 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</span><span class="cit">2020 Apr 8;20(4):2296-2302.</span> </div> <span class="citation-doi"> doi: 10.1021/acs.nanolett.9b04561. </span> <span class="secondary-date"> Epub 2020 Mar 12. </span> </div> <h1 class="heading-title"> Multiplexed Continuous Biosensing by Single-Molecule Encoded Nanoswitches </h1> <div class="inline-authors"> <div class="authors"> <div class="authors-list"> <span class="authors-list-item "><a class="full-name" href="/?term=Lubken+RM&cauthor_id=32091908" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="Rafiq M Lubken">Rafiq M Lubken</a><sup class="affiliation-links"><span class="author-sup-separator"> </span><a class="affiliation-link" title="Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands." href="#full-view-affiliation-1" ref="linksrc=author_aff"> 1 </a><span class="author-sup-separator"> </span><a class="affiliation-link" title="Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands." href="#full-view-affiliation-2" ref="linksrc=author_aff"> 2 </a></sup><span class="comma">, </span></span><span class="authors-list-item "><a class="full-name" href="/?term=de+Jong+AM&cauthor_id=32091908" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="Arthur M de Jong">Arthur M de Jong</a><sup class="affiliation-links"><span class="author-sup-separator"> </span><a class="affiliation-link" title="Department of Applied Physics, Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands." href="#full-view-affiliation-3" ref="linksrc=author_aff"> 3 </a><span class="author-sup-separator"> </span><a class="affiliation-link" title="Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands." href="#full-view-affiliation-2" ref="linksrc=author_aff"> 2 </a></sup><span class="comma">, </span></span><span class="authors-list-item "><a class="full-name" href="/?term=Prins+MWJ&cauthor_id=32091908" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="Menno W J Prins">Menno W J Prins</a><sup class="affiliation-links"><span class="author-sup-separator"> </span><a class="affiliation-link" title="Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands." href="#full-view-affiliation-1" ref="linksrc=author_aff"> 1 </a><span class="author-sup-separator"> </span><a class="affiliation-link" title="Department of Applied Physics, Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands." href="#full-view-affiliation-3" ref="linksrc=author_aff"> 3 </a><span class="author-sup-separator"> </span><a class="affiliation-link" title="Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands." href="#full-view-affiliation-2" ref="linksrc=author_aff"> 2 </a></sup></span> </div> </div> </div> <div class="short-article-details"> Affiliations <button class="more-details" id="toggle-authors" data-alt-text="Collapse" aria-controls="expanded-authors" aria-expanded="false" data-pinger-ignore data-fetch-url="/32091908/long-authors/" data-id-prefix=""> <span class="title"> Expand </span> </button> </div> <div class="extended-article-details" id="expanded-authors" aria-hidden="true"> <div class="expanded-authors" id="full-view-expanded-authors"> <div class="affiliations"> <h3 class="title"> Affiliations </h3> <ul class="item-list"> <li data-affiliation-id="full-view-affiliation-1" id="full-view-affiliation-1" ><sup class="key">1</sup> Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands.</li> <li data-affiliation-id="full-view-affiliation-2" id="full-view-affiliation-2" ><sup class="key">2</sup> Institute for Complex 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</span> </div> <div class="expanded-authors" id="short-view-expanded-authors"> <div class="authors"> <h3 class="title"> Authors </h3> <div class="authors-list"> <span class="authors-list-item "><a class="full-name" href="/?term=Lubken+RM&cauthor_id=32091908" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="Rafiq M Lubken">Rafiq M Lubken</a><sup class="affiliation-links"><span class="author-sup-separator"> </span><a class="affiliation-link" title="Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands." href="#short-view-affiliation-1" ref="linksrc=author_aff"> 1 </a><span class="author-sup-separator"> </span><a class="affiliation-link" title="Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands." href="#short-view-affiliation-2" ref="linksrc=author_aff"> 2 </a></sup><span class="comma">, </span></span><span class="authors-list-item "><a class="full-name" href="/?term=de+Jong+AM&cauthor_id=32091908" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="Arthur M de Jong">Arthur M de Jong</a><sup class="affiliation-links"><span class="author-sup-separator"> </span><a class="affiliation-link" title="Department of Applied Physics, Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands." href="#short-view-affiliation-3" ref="linksrc=author_aff"> 3 </a><span class="author-sup-separator"> </span><a class="affiliation-link" title="Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands." href="#short-view-affiliation-2" ref="linksrc=author_aff"> 2 </a></sup><span class="comma">, </span></span><span class="authors-list-item "><a class="full-name" href="/?term=Prins+MWJ&cauthor_id=32091908" ref="linksrc=author_name_link" data-ga-category="search" data-ga-action="author_link" data-ga-label="Menno W J Prins">Menno W J Prins</a><sup class="affiliation-links"><span class="author-sup-separator"> </span><a class="affiliation-link" title="Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands." href="#short-view-affiliation-1" ref="linksrc=author_aff"> 1 </a><span class="author-sup-separator"> </span><a class="affiliation-link" title="Department of Applied Physics, Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands." href="#short-view-affiliation-3" ref="linksrc=author_aff"> 3 </a><span class="author-sup-separator"> </span><a class="affiliation-link" title="Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands." href="#short-view-affiliation-2" ref="linksrc=author_aff"> 2 </a></sup></span> </div> </div> <div class="affiliations"> <h3 class="title"> Affiliations </h3> <ul class="item-list"> <li data-affiliation-id="short-view-affiliation-1" id="short-view-affiliation-1" ><sup class="key">1</sup> Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands.</li> <li data-affiliation-id="short-view-affiliation-2" id="short-view-affiliation-2" ><sup class="key">2</sup> Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands.</li> <li data-affiliation-id="short-view-affiliation-3" id="short-view-affiliation-3" ><sup class="key">3</sup> Department of Applied Physics, Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands.</li> </ul> </div> </div> <ul class="identifiers" id="short-view-identifiers"> <li> <span class="identifier pubmed"> <span class="id-label"> PMID: </span> <strong class="current-id" title="PubMed ID">32091908</strong> </span> </li> <li> <span class="identifier pmc"> <span class="id-label"> PMCID: </span> <a class="id-link" target="_blank" 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different styles" data-ga-category="save_share" data-ga-action="cite" data-ga-label="open" data-all-citations-url="/32091908/citations/" data-citation-style="nlm" data-pubmed-format-link="/32091908/export/"><span class="button-label">Cite</span></button><button class="more-actions-button more-actions-dialog-trigger" title="Open dialog with more actions to take" ref="linksrc=more_actions_btn"></button><div class="display-options" data-format-key="format" data-default-format-value="abstract" data-current-format-value="abstract" role="region"><button class="trigger usa-button" title="Change format" data-ga-category="display_options" data-ga-action="display_options_dialog"><span class="button-label">Display options</span></button><div class="dropdown" hidden><div class="title"> Display options </div><div class="content"><div class="format-container"><span class="option-label"> Format </span><select name="format" data-initial-value="abstract" aria-label="Display format" id="inline-actions-display-format"><option value="abstract">Abstract</option><option value="pubmed">PubMed</option><option value="pmid">PMID</option></select></div></div></div></div></div></div> <div class="abstract" id="abstract"> <h2 class="title"> Abstract </h2> <div class="abstract-content selected" id="eng-abstract"> <p> Single-molecule techniques have become impactful in bioanalytical sciences, though the advantages for continuous biosensing are yet to be discovered. Here we present a multiplexed, continuous biosensing method, enabled by an analyte-sensitive, single-molecular nanoswitch with a particle as a reporter. The nanoswitch opens and closes under the influence of single target molecules. This reversible switching yields binary transitions between two highly reproducible states, enabling reliable quantification of the single-molecule kinetics. The multiplexing functionality is encoded per particle via the dissociation characteristics of the nanoswitch, while the target concentration is revealed by the association characteristics. We demonstrate by experiments and simulations the multiplexed, continuous monitoring of oligonucleotide targets, at picomolar concentrations in buffer and in filtered human blood plasma. </p> </div> <p> <strong class="sub-title"> Keywords: </strong> continuous biosensing; kinetic identification; multiplexing; single-molecule biosensing. </p> </div> <p class="disclaimer" id="disclaimer"> <a href="/disclaimer/" target="_blank" ga_category="literature_resources" ga_action="disclaimer_link" ga_label="under_abstract">PubMed Disclaimer</a> </p> <div class="conflict-of-interest" id="conflict-of-interest"> <h2 class="title"> Conflict of interest statement </h2> <div class="statement"> <p xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:p1="http://pubmed.gov/pub-one">The authors declare the following competing financial interest(s): M.W.J.P. is listed as one of the inventors on patent application WO/2016/096901 (Biosensor based on a tethered particle). M.W.J.P., A.M.d.J., and R.M.L. are listed as inventors on a filed patent application relating to the data presented in this paper. M.W.J.P. is cofounder of Helia Biomonitoring BV that has a license to these patents. All authors declare no further competing interests.</p> </div> </div> <div class="figures" id="figures"> <h2 class="title"> Figures </h2> <div class="figures-list" id="slides-container" itemscope itemtype="http://schema.org/ImageGallery"> <figure class="figure-item " itemscope itemtype="http://schema.org/ImageObject" itemprop="associatedMedia" data-slide-index="0" data-label-slug="figure-1"> <a class="figure-link" href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a602/7252944/6e8c90f2112c/nl9b04561_0001.jpg" itemprop="contentUrl" aria-describedby="figure-caption-0" role="button" data-image-width="705" data-image-height="644" data-image-alt="Figure 1" data-pmc-id="PMC7252944" data-figure-id="fig1"> <img class="figure-thumb" itemprop="thumbnail" id="article-image-0" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a602/7252944/99e51714d6dc/nl9b04561_0001.gif" alt="Figure 1" /> </a> <meta itemprop="width" itemtype="http://schema.org/ImageObject" content="705"> <meta itemprop="height" itemtype="http://schema.org/ImageObject" content="644"> <div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true"> <div class="caption-wrap"> <strong class="figure-label"> <p> Figure 1 </p> </strong> <div class="figure-caption-contents"><p> Sensor concept with a single-molecule… </p></div> </div> </div> <div class="figure-caption-medium figure-caption-text" aria-hidden="true"> <strong class="figure-label"> <p> Figure 1 </p> </strong> <div class="figure-caption-contents"><p> Sensor concept with a single-molecule encoded binary nanoswitch for multiplexed continuous biomolecular monitoring.… </p></div> </div> <figcaption id="figure-caption-0" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description"> <strong class="figure-label"> Figure 1 </strong> <div class="figure-caption-contents">Sensor concept with a single-molecule encoded binary nanoswitch for multiplexed continuous biomolecular monitoring. (a) Micrometer-sized particles (yellow) are tethered to a substrate using a 56 nm dsDNA stem (black). The particle is functionalized with particle probes of type <i>i</i> (dark green) and a single stem probe of type <i>i</i> (green). Both probes bind reversibly to a single target molecule of type <i>i</i> (light green) present in solution. The inset shows schematically the DNA-based nanoswitch conjugated to the particle by a NeutrAvidin–biotin interaction and to the substrate by an antibody–antigen (digoxigenin) interaction. A detailed overview of the DNA sequences is given in the <ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="uri" xlink:href="http://pubs.acs.org/doi/suppl/10.1021/acs.nanolett.9b04561/suppl_file/nl9b04561_si_001.pdf">Supporting Information</ext-link>. (b) Target molecule binding to the nanoswitch causes the particle to exhibit either of two concentric motion patterns corresponding to the unbound (high mobility) and bound state (low mobility). (c) The radial position of a particle over time shows binary transitions caused by single-molecule binding and unbinding events. The distribution of observed bound state lifetimes per particle can be used to distinguish between, e.g., low-affinity (particles <i>i</i>, blue) and high-affinity (particles <i>j</i>, red) target-specific particle subpopulations, referred to as kinetic identification. Examples of raw data traces are shown in the <ext-link xmlns:xlink="http://www.w3.org/1999/xlink" ext-link-type="uri" xlink:href="http://pubs.acs.org/doi/suppl/10.1021/acs.nanolett.9b04561/suppl_file/nl9b04561_si_001.pdf">Supporting Information</ext-link>. (d) Hundreds of particles, each functionalized with an encoded binary nanoswitch, are observed simultaneously. By kinetic identification based on the dissociation kinetics, each particle can be assigned to a target-specific particle subpopulation. For each particle subpopulation, the respective target concentration can be determined over time using the measured association kinetics.</div> </figcaption> </figure> <figure class="figure-item " itemscope itemtype="http://schema.org/ImageObject" itemprop="associatedMedia" data-slide-index="1" data-label-slug="figure-2"> <a class="figure-link" href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a602/7252944/73b715f815ad/nl9b04561_0002.jpg" itemprop="contentUrl" aria-describedby="figure-caption-1" role="button" data-image-width="697" data-image-height="830" data-image-alt="Figure 2" data-pmc-id="PMC7252944" data-figure-id="fig2"> <img class="figure-thumb" itemprop="thumbnail" id="article-image-1" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a602/7252944/20688fb0a5d5/nl9b04561_0002.gif" alt="Figure 2" /> </a> <meta itemprop="width" itemtype="http://schema.org/ImageObject" content="697"> <meta itemprop="height" itemtype="http://schema.org/ImageObject" content="830"> <div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true"> <div class="caption-wrap"> <strong class="figure-label"> <p> Figure 2 </p> </strong> <div class="figure-caption-contents"><p> Performance of continuous biomarker monitoring… </p></div> </div> </div> <div class="figure-caption-medium figure-caption-text" aria-hidden="true"> <strong class="figure-label"> <p> Figure 2 </p> </strong> <div class="figure-caption-contents"><p> Performance of continuous biomarker monitoring in buffer and blood plasma. (a, b) Mid-affinity… </p></div> </div> <figcaption id="figure-caption-1" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description"> <strong class="figure-label"> Figure 2 </strong> <div class="figure-caption-contents">Performance of continuous biomarker monitoring in buffer and blood plasma. (a, b) Mid-affinity target concentration dependencies of the bound (red) and unbound state lifetime (blue). The data shows that <i>τ</i><sub>B</sub> = 15.2 ± 0.3 s and <i>τ</i><sub>B</sub> = 12.5 ± 0.5 s and that <i>τ</i><sub>U</sub> scales approximately as <i>τ</i><sub>U</sub> ∝ [<i>T</i>]<sup>−0.8±0.1</sup> and [<i>T</i>]<sup>−0.64±0.09</sup> for buffer and blood plasma conditions, respectively. (c) Dose–response curves of DNA targets in buffer, with a high, mid, and low affinity to the particle probe. Hill equation fits (solid lines) yield EC<sub>50</sub> values of 14 ± 6 pM, 0.17 ± 0.05 nM, and 1.7 ± 0.3 nM, respectively. (d) Dose–response curve in blood plasma of the mid-affinity DNA target. Hill equation fit (solid line) yields EC<sub>50</sub> of 0.4 ± 0.1 nM. The inset shows the data on a double linear scale. (e–g) Concentration response traces for low- and mid-affinity DNA targets in buffer and mid-affinity DNA target in blood plasma. The respective relaxation times are <i>τ</i><sub>R</sub> = 10 ± 1 min, <i>τ</i><sub>R</sub> = 41 ± 6 min, and <i>τ</i><sub>R</sub> = 37 ± 8 min, resulting from the single-exponential fits (red). Reported errors are the standard errors of the fit. The error bars in the activity graphs are the stochastic errors and are mostly smaller than the symbol size. The shading in panels c and d indicates the 95% confidence interval of the Hill equation fit. The number of particles per data point was between 15 and 100, measured in two microscopic fields of view.</div> </figcaption> </figure> <figure class="figure-item " itemscope itemtype="http://schema.org/ImageObject" itemprop="associatedMedia" data-slide-index="2" data-label-slug="figure-3"> <a class="figure-link" href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a602/7252944/19d5ab120dfd/nl9b04561_0003.jpg" itemprop="contentUrl" aria-describedby="figure-caption-2" role="button" data-image-width="686" data-image-height="560" data-image-alt="Figure 3" data-pmc-id="PMC7252944" data-figure-id="fig3"> <img class="figure-thumb" itemprop="thumbnail" id="article-image-2" src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a602/7252944/fd520e9bb17f/nl9b04561_0003.gif" alt="Figure 3" /> </a> <meta itemprop="width" itemtype="http://schema.org/ImageObject" content="686"> <meta itemprop="height" itemtype="http://schema.org/ImageObject" content="560"> <div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true"> <div class="caption-wrap"> <strong class="figure-label"> <p> Figure 3 </p> </strong> <div class="figure-caption-contents"><p> Multiplexing performance by kinetic identification… </p></div> </div> </div> <div class="figure-caption-medium figure-caption-text" aria-hidden="true"> <strong class="figure-label"> <p> Figure 3 </p> </strong> <div class="figure-caption-contents"><p> Multiplexing performance by kinetic identification of nanoswitches. (a) Concentration dependencies of the bound… </p></div> </div> <figcaption id="figure-caption-2" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description"> <strong class="figure-label"> Figure 3 </strong> <div class="figure-caption-contents">Multiplexing performance by kinetic identification of nanoswitches. (a) Concentration dependencies of the bound state lifetime for low- and high-affinity targets (blue and red, respectively). The data indicate that <i>τ</i><sub>B</sub> = 15.4 ± 0.1 s and <i>τ</i><sub>B</sub> = 86 ± 2 s for low- and high-affinity targets, respectively. (b) Bound state lifetime distribution per particle for low- and high-affinity targets. The threshold and window width to discard ambiguous particles used for kinetic identification are indicated by the black line and shaded area, respectively. The dashed black lines show log-normal distributions. The inset shows simulated bound state lifetime distributions for both affinities, for a 10 min (solid lines) and 30 min (dashed lines) measurement. (c) Receiver operating curve that quantifies the performance of the kinetic identification. An optimum with a kinetic sensitivity of 97% and a kinetic specificity of 88% was found using a zero window width; see the red dot. The inset shows the potential of discarding ambiguous particles, by the approximate trends of the kinetic sensitivity and specificity as a function of the window width. (d) Control experiment to quantify the cross-talk between particle populations in the sensor. Flow cell A contains particles specific for target A and flow cell B for target B. The concentration–time profiles show how the targets are applied to each individual flow cell. Both sensors only respond to their specific target. (e) CDF of the bound state lifetimes with τ<sub>B</sub><sup>1</sup> = 13.7 ± 0.1 s and τ<sub>B</sub><sup arrange="stagger">2</sup> = 113 ± 2 s resulting from a double-exponential fit (yellow). The inset shows the CDFs of the two separate particle populations. The single-exponential fits (yellow) give <i>τ</i><sub>B</sub> = 14.4 ± 0.1 s and <i>τ</i><sub>B</sub> = 113 ± 2 s for the low- and high-affinity interaction, respectively. (f) Simulated bound state lifetime distributions for a 30 min measurement show the multiplexing potential with the current experimental limits. The blue and red distributions have mean bound state lifetimes that are matched with the lifetimes found in panel e. Reported errors are the standard errors of the fit. The number of particles per data point was between 15 and 100.</div> </figcaption> </figure> </div>  <div class="pswp figures-dialog" id="figures-dialog" tabindex="-1" role="dialog" aria-hidden="true" data-image-gallery-uid="article-figures" aria-label="Figures Dialog">  <div class="pswp__bg"></div>  <div class="pswp__scroll-wrap">  <div class="pswp__container"> <div class="pswp__item"></div> <div class="pswp__item"></div> <div class="pswp__item"></div> </div>  <div class="pswp__ui pswp__ui--hidden"> <div class="pswp__top-bar">  <div class="pswp__counter"></div> <button class="pswp__button pswp__button--close" title="Close (Esc)"></button> <button class="pswp__button pswp__button--share" title="Share"></button> <button class="pswp__button pswp__button--fs" title="Toggle fullscreen"></button> <button class="pswp__button pswp__button--zoom" title="Zoom in/out"></button>   <div class="pswp__preloader"> <div class="pswp__preloader__icn"> <div class="pswp__preloader__cut"> <div class="pswp__preloader__donut"></div> </div> </div> </div> </div> <div class="pswp__share-modal pswp__share-modal--hidden pswp__single-tap"> <div class="pswp__share-tooltip"></div> </div> <button class="pswp__button pswp__button--arrow--left" title="Previous (arrow left)"> </button> <button class="pswp__button pswp__button--arrow--right" title="Next (arrow right)"> </button> <div class="pswp__caption"> <div class="pswp__caption__center"></div> </div> </div> </div> </div> <div class="figure-dialog-caption"> <div class="figure-caption-link-container"> <a class="figure-caption-link" href="#">See this image and copyright information in PMC</a> </div> <div class="figure-caption-center" data-expand-title="click to show full caption" data-collapse-title="click to hide full caption"> </div> <button class="figure-caption-toggle-btn" aria-label="show full caption" data-expand-text="show full caption" data-collapse-text="hide full caption">  </button> </div> </div> <div class="similar-articles" id="similar"> <h2 class="title"> Similar articles </h2> <ul class="articles-list" id="similar-articles-list"> <li class="full-docsum"> <div class="docsum-content" style="display:inline;"> <a class="docsum-title" href="/37220424/" ref="article_id=37220424&linksrc=similar_articles_link&ordinalpos=1" data-ga-category="similar_article" data-ga-action="37220424" data-ga-label=""> Multiplexed Nanopore-Based Nucleic Acid Sensing and Bacterial Identification Using DNA Dumbbell Nanoswitches. </a> <div class="docsum-citation full-citation"> <span class="docsum-authors full-authors">Zhu J, Tivony R, Bošković F, Pereira-Dias J, Sandler SE, Baker S, Keyser UF.</span> <span class="docsum-authors short-authors">Zhu J, et al.</span> <span class="docsum-journal-citation full-journal-citation">J Am Chem Soc. 2023 Jun 7;145(22):12115-12123. doi: 10.1021/jacs.3c01649. 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Biotechnol. 2019, 37, 407–419. 10.1038/s41587-019-0040-3. - <a class="reference-link" href="https://doi.org/10.1038/s41587-019-0040-3" ref="linksrc=references_link&ordinalpos=1" data-ga-category="reference" data-ga-action="10.1038/s41587-019-0040-3" > DOI </a> - <a class="reference-link" href="/30804536/" ref="linksrc=references_link&ordinalpos=2" data-ga-category="reference" data-ga-action="30804536" > PubMed </a> </li> </ol> </li> </ol> <div class="actions-bar"> <button class="show-all" ref="linksrc=show_all_references_link" aria-controls="top-references-list-1" data-next-page-url="/32091908/references/" data-ga-category="reference" data-ga-action="show_more" > Show all 14 references </button> </div> </div> </div> </div> <div id="mesh-terms" class="mesh-terms keywords-section"> <h2 class="title"> MeSH terms </h2> <ul class="keywords-list"><li><div class="keyword-actions dropdown-block"><button class="keyword-actions-trigger trigger keyword-link major" aria-label="Toggle dropdown menu 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Multiplexed Continuous Biosensing by Single-Molecule Encoded Nanoswitches - PubMed