Bioluminescent RAPPID Sensors for the Single-Step Detection of Soluble Axl and Multiplex Analysis of Cell Surface Cancer Biomarkers
Early diagnosis of cancer is essential for the efficacy of treatment. Our group recently developed RAPPID, a bioluminescent immunoassay platform capable of measuring a wide panel of biomarkers directly in solution. Here, we developed and systematically screened different RAPPID sensors for sensitive...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2022-05, Vol.94 (17), p.6548-6556 |
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| creator | van Aalen, Eva A Wouters, Simone F. A Verzijl, Dennis Merkx, Maarten |
| description | Early diagnosis of cancer is essential for the efficacy of treatment. Our group recently developed RAPPID, a bioluminescent immunoassay platform capable of measuring a wide panel of biomarkers directly in solution. Here, we developed and systematically screened different RAPPID sensors for sensitive detection of the soluble fraction of Axl (sAxl), a cell surface receptor that is overexpressed in several types of cancer. The best-performing RAPPID sensor, with a limit of detection of 8 pM and a >9-fold maximal change in emission ratio, was applied to measure Axl in three different contexts: clinically relevant sAxl levels (∼0.5 and ∼1 nM) in diluted blood plasma, proteolytically cleaved Axl in the cell culture medium of A431 and HeLa cancer cells, and Axl on the membrane of A431 cells. We further extended the sensor toolbox by developing dual-color RAPPID for simultaneous detection of Axl and EGFR on A431 and HeLa cells, as well as an AND-gate RAPPID that measures the concurrent presence of these two cell surface receptors on the same cell. These new RAPPID sensors provide attractive alternatives for more laborious protein detection and quantification methods such as FACS and immunostainings, due to their simple practical implantation and low intrinsic background signal. |
| doi_str_mv | 10.1021/acs.analchem.2c00297 |
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A ; Verzijl, Dennis ; Merkx, Maarten</creator><creatorcontrib>van Aalen, Eva A ; Wouters, Simone F. A ; Verzijl, Dennis ; Merkx, Maarten</creatorcontrib><description>Early diagnosis of cancer is essential for the efficacy of treatment. Our group recently developed RAPPID, a bioluminescent immunoassay platform capable of measuring a wide panel of biomarkers directly in solution. Here, we developed and systematically screened different RAPPID sensors for sensitive detection of the soluble fraction of Axl (sAxl), a cell surface receptor that is overexpressed in several types of cancer. The best-performing RAPPID sensor, with a limit of detection of 8 pM and a >9-fold maximal change in emission ratio, was applied to measure Axl in three different contexts: clinically relevant sAxl levels (∼0.5 and ∼1 nM) in diluted blood plasma, proteolytically cleaved Axl in the cell culture medium of A431 and HeLa cancer cells, and Axl on the membrane of A431 cells. We further extended the sensor toolbox by developing dual-color RAPPID for simultaneous detection of Axl and EGFR on A431 and HeLa cells, as well as an AND-gate RAPPID that measures the concurrent presence of these two cell surface receptors on the same cell. These new RAPPID sensors provide attractive alternatives for more laborious protein detection and quantification methods such as FACS and immunostainings, due to their simple practical implantation and low intrinsic background signal.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/acs.analchem.2c00297</identifier><identifier>PMID: 35438976</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Analytical chemistry ; Axl protein ; Bioluminescence ; Biomarkers ; Biomarkers, Tumor ; Blood plasma ; Cancer ; Cell culture ; Cell surface ; Chemistry ; Early Detection of Cancer ; Emission measurements ; Flow cytometry ; HeLa Cells ; Humans ; Immunoassay ; Intercellular Signaling Peptides and Proteins ; Neoplasms ; Proto-Oncogene Proteins - metabolism ; Receptor Protein-Tyrosine Kinases - metabolism ; Receptors ; Sensors</subject><ispartof>Analytical chemistry (Washington), 2022-05, Vol.94 (17), p.6548-6556</ispartof><rights>2022 The Authors. Published by American Chemical Society</rights><rights>Copyright American Chemical Society May 3, 2022</rights><rights>2022 The Authors. Published by American Chemical Society 2022 The Authors</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a4587-231f0f8f11e0603467db04b728455f1622cec3f57d60b009444bab209caaf33c3</citedby><cites>FETCH-LOGICAL-a4587-231f0f8f11e0603467db04b728455f1622cec3f57d60b009444bab209caaf33c3</cites><orcidid>0000-0001-9484-3882</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/acs.analchem.2c00297$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/acs.analchem.2c00297$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>230,314,776,780,881,2752,27053,27901,27902,56713,56763</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35438976$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>van Aalen, Eva A</creatorcontrib><creatorcontrib>Wouters, Simone F. A</creatorcontrib><creatorcontrib>Verzijl, Dennis</creatorcontrib><creatorcontrib>Merkx, Maarten</creatorcontrib><title>Bioluminescent RAPPID Sensors for the Single-Step Detection of Soluble Axl and Multiplex Analysis of Cell Surface Cancer Biomarkers</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>Early diagnosis of cancer is essential for the efficacy of treatment. Our group recently developed RAPPID, a bioluminescent immunoassay platform capable of measuring a wide panel of biomarkers directly in solution. Here, we developed and systematically screened different RAPPID sensors for sensitive detection of the soluble fraction of Axl (sAxl), a cell surface receptor that is overexpressed in several types of cancer. The best-performing RAPPID sensor, with a limit of detection of 8 pM and a >9-fold maximal change in emission ratio, was applied to measure Axl in three different contexts: clinically relevant sAxl levels (∼0.5 and ∼1 nM) in diluted blood plasma, proteolytically cleaved Axl in the cell culture medium of A431 and HeLa cancer cells, and Axl on the membrane of A431 cells. We further extended the sensor toolbox by developing dual-color RAPPID for simultaneous detection of Axl and EGFR on A431 and HeLa cells, as well as an AND-gate RAPPID that measures the concurrent presence of these two cell surface receptors on the same cell. These new RAPPID sensors provide attractive alternatives for more laborious protein detection and quantification methods such as FACS and immunostainings, due to their simple practical implantation and low intrinsic background signal.</description><subject>Analytical chemistry</subject><subject>Axl protein</subject><subject>Bioluminescence</subject><subject>Biomarkers</subject><subject>Biomarkers, Tumor</subject><subject>Blood plasma</subject><subject>Cancer</subject><subject>Cell culture</subject><subject>Cell surface</subject><subject>Chemistry</subject><subject>Early Detection of Cancer</subject><subject>Emission measurements</subject><subject>Flow cytometry</subject><subject>HeLa Cells</subject><subject>Humans</subject><subject>Immunoassay</subject><subject>Intercellular Signaling Peptides and Proteins</subject><subject>Neoplasms</subject><subject>Proto-Oncogene Proteins - metabolism</subject><subject>Receptor Protein-Tyrosine Kinases - metabolism</subject><subject>Receptors</subject><subject>Sensors</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kc1u1DAUhS0EokPhDRCyxIZNhmsncZIN0jClUKmIisDacjzXHRcnHuwEteu-OI5mOgIWLKy78HfO_TmEvGSwZMDZW6XjUg3K6S32S64BeFM9IgtWcshEXfPHZAEAecYrgBPyLMYbAMaAiafkJC-LvG4qsSD37613U28HjBqHkX5dXV1dnNEWh-hDpMYHOm6Rtna4dpi1I-7oGY6oR-sH6g1tk7pzSFe3jqphQz9PbrQ7h7d0lWa7izbO1Bqdo-0UjNJI12rQGGhq3KvwA0N8Tp4Y5SK-ONRT8v38w7f1p-zyy8eL9eoyU0VZVxnPmQFTG8YQBOSFqDYdFF3F66IsDROca9S5KauNgA6gKYqiUx2HRitl8lznp-Td3nc3dT1u5n2DcnIXbBrkTnpl5d8_g93Ka_9LNiCadLBk8OZgEPzPCeMoe5vO5pwa0E9RclHyOj3RJPT1P-iNn0I6yUwJYDytUCWq2FM6-BgDmuMwDOScskwpy4eU5SHlJHv15yJH0UOsCYA9MMuPjf_r-RtHJ7eF</recordid><startdate>20220503</startdate><enddate>20220503</enddate><creator>van Aalen, Eva A</creator><creator>Wouters, Simone F. 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A</creatorcontrib><creatorcontrib>Verzijl, Dennis</creatorcontrib><creatorcontrib>Merkx, Maarten</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>van Aalen, Eva A</au><au>Wouters, Simone F. A</au><au>Verzijl, Dennis</au><au>Merkx, Maarten</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Bioluminescent RAPPID Sensors for the Single-Step Detection of Soluble Axl and Multiplex Analysis of Cell Surface Cancer Biomarkers</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2022-05-03</date><risdate>2022</risdate><volume>94</volume><issue>17</issue><spage>6548</spage><epage>6556</epage><pages>6548-6556</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><abstract>Early diagnosis of cancer is essential for the efficacy of treatment. Our group recently developed RAPPID, a bioluminescent immunoassay platform capable of measuring a wide panel of biomarkers directly in solution. Here, we developed and systematically screened different RAPPID sensors for sensitive detection of the soluble fraction of Axl (sAxl), a cell surface receptor that is overexpressed in several types of cancer. The best-performing RAPPID sensor, with a limit of detection of 8 pM and a >9-fold maximal change in emission ratio, was applied to measure Axl in three different contexts: clinically relevant sAxl levels (∼0.5 and ∼1 nM) in diluted blood plasma, proteolytically cleaved Axl in the cell culture medium of A431 and HeLa cancer cells, and Axl on the membrane of A431 cells. We further extended the sensor toolbox by developing dual-color RAPPID for simultaneous detection of Axl and EGFR on A431 and HeLa cells, as well as an AND-gate RAPPID that measures the concurrent presence of these two cell surface receptors on the same cell. These new RAPPID sensors provide attractive alternatives for more laborious protein detection and quantification methods such as FACS and immunostainings, due to their simple practical implantation and low intrinsic background signal.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>35438976</pmid><doi>10.1021/acs.analchem.2c00297</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0001-9484-3882</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Analytical chemistry Axl protein Bioluminescence Biomarkers Biomarkers, Tumor Blood plasma Cancer Cell culture Cell surface Chemistry Early Detection of Cancer Emission measurements Flow cytometry HeLa Cells Humans Immunoassay Intercellular Signaling Peptides and Proteins Neoplasms Proto-Oncogene Proteins - metabolism Receptor Protein-Tyrosine Kinases - metabolism Receptors Sensors |
| title | Bioluminescent RAPPID Sensors for the Single-Step Detection of Soluble Axl and Multiplex Analysis of Cell Surface Cancer Biomarkers |
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