Microfluidic-Based Detection of AML-Specific Biomarkers Using the Example of Promyelocyte Leukemia
A microfluidic assay for the detection of promyelocytic leukemia (PML)-retinoic acid receptor α (RARα) fusion protein was developed. This microfluidic-based system can be used for rapid personalized differential diagnosis of acute promyelocyte leukemia (APL) with the aim of early initiation of indiv...
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| Veröffentlicht in: | International journal of molecular sciences 2020-11, Vol.21 (23), p.8942 |
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| description | A microfluidic assay for the detection of promyelocytic leukemia (PML)-retinoic acid receptor α (RARα) fusion protein was developed. This microfluidic-based system can be used for rapid personalized differential diagnosis of acute promyelocyte leukemia (APL) with the aim of early initiation of individualized therapy. The fusion protein PML-RARα occurs in 95% of acute promyelocytic leukemia cases and is considered as diagnostically relevant. The fusion protein is formed as a result of translocation t(15,17) and is detected in the laboratory by fluorescence in situ hybridization (FISH) or reverse transcriptase polymerase chain reaction (RT-PCR). Diagnostic methods require many laboratory steps with specialized staff. The developed microfluidic assay includes a sandwich enzyme-linked immunosorbent assay (ELISA) system for PML-RARα on surface of magnetic microparticles in a microfluidic chip. A rapid detection of PML-RARα in cell lysates is achieved in less than one hour. A biotinylated PML-antibody on the surface of magnetic streptavidin coated microparticles is used as capture antibody. The bound translocation product is detected by a RARα antibody conjugated with horseradish peroxidase and the substrate QuantaRed. The analysis is performed in microfluidic channels which involves automated liquid processing with stringent washing and short incubation times. The results of the developed assay show that cell lysates of PML-RARα-positive cells (NB-4) can be clearly distinguished from PML-RARα-negative cells (HL-60, MV4-11). |
| doi_str_mv | 10.3390/ijms21238942 |
| format | Article |
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This microfluidic-based system can be used for rapid personalized differential diagnosis of acute promyelocyte leukemia (APL) with the aim of early initiation of individualized therapy. The fusion protein PML-RARα occurs in 95% of acute promyelocytic leukemia cases and is considered as diagnostically relevant. The fusion protein is formed as a result of translocation t(15,17) and is detected in the laboratory by fluorescence in situ hybridization (FISH) or reverse transcriptase polymerase chain reaction (RT-PCR). Diagnostic methods require many laboratory steps with specialized staff. The developed microfluidic assay includes a sandwich enzyme-linked immunosorbent assay (ELISA) system for PML-RARα on surface of magnetic microparticles in a microfluidic chip. A rapid detection of PML-RARα in cell lysates is achieved in less than one hour. A biotinylated PML-antibody on the surface of magnetic streptavidin coated microparticles is used as capture antibody. The bound translocation product is detected by a RARα antibody conjugated with horseradish peroxidase and the substrate QuantaRed. The analysis is performed in microfluidic channels which involves automated liquid processing with stringent washing and short incubation times. The results of the developed assay show that cell lysates of PML-RARα-positive cells (NB-4) can be clearly distinguished from PML-RARα-negative cells (HL-60, MV4-11).</description><identifier>ISSN: 1422-0067</identifier><identifier>ISSN: 1661-6596</identifier><identifier>EISSN: 1422-0067</identifier><identifier>DOI: 10.3390/ijms21238942</identifier><identifier>PMID: 33255664</identifier><language>eng</language><publisher>Switzerland: MDPI AG</publisher><subject>Acute myeloid leukemia ; Acute promyeloid leukemia ; Antibodies ; Antigens ; Assaying ; Automation ; Biomarkers, Tumor - genetics ; Biomarkers, Tumor - isolation & purification ; Chromosomes ; Diagnostic systems ; Differential diagnosis ; Enzyme-linked immunosorbent assay ; Enzymes ; Fluorescence ; Fluorescence in situ hybridization ; Fusion protein ; Genes ; Granulocyte Precursor Cells - metabolism ; Granulocyte Precursor Cells - pathology ; Horseradish peroxidase ; Humans ; In Situ Hybridization, Fluorescence - methods ; Lab-on-a-chip ; Laboratories ; Leukemia ; Leukemia, Promyelocytic, Acute - diagnosis ; Leukemia, Promyelocytic, Acute - genetics ; Leukemia, Promyelocytic, Acute - pathology ; Lysates ; Microfluidics ; Microfluidics - methods ; Oncogene Proteins, Fusion - genetics ; Oncogene Proteins, Fusion - isolation & purification ; Patients ; Peroxidase ; Point of care testing ; Polymerase chain reaction ; Precision Medicine ; Promyelocytic Leukemia Protein - genetics ; Promyeloid leukemia ; Proteins ; Retinoic acid ; Retinoic Acid Receptor alpha - genetics ; RNA-directed DNA polymerase ; Streptavidin ; Translocation, Genetic - genetics</subject><ispartof>International journal of molecular sciences, 2020-11, Vol.21 (23), p.8942</ispartof><rights>2020. This work is licensed under http://creativecommons.org/licenses/by/3.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2020 by the authors. 2020</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c412t-590e7f781d8238fec536b09fb9276d3b7d9f57e612a1d60471baf57ba06c8b853</citedby><cites>FETCH-LOGICAL-c412t-590e7f781d8238fec536b09fb9276d3b7d9f57e612a1d60471baf57ba06c8b853</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC7728129/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC7728129/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,881,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33255664$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Emde, Benedikt</creatorcontrib><creatorcontrib>Kreher, Heike</creatorcontrib><creatorcontrib>Bäumer, Nicole</creatorcontrib><creatorcontrib>Bäumer, Sebastian</creatorcontrib><creatorcontrib>Bouwes, Dominique</creatorcontrib><creatorcontrib>Tickenbrock, Lara</creatorcontrib><title>Microfluidic-Based Detection of AML-Specific Biomarkers Using the Example of Promyelocyte Leukemia</title><title>International journal of molecular sciences</title><addtitle>Int J Mol Sci</addtitle><description>A microfluidic assay for the detection of promyelocytic leukemia (PML)-retinoic acid receptor α (RARα) fusion protein was developed. This microfluidic-based system can be used for rapid personalized differential diagnosis of acute promyelocyte leukemia (APL) with the aim of early initiation of individualized therapy. The fusion protein PML-RARα occurs in 95% of acute promyelocytic leukemia cases and is considered as diagnostically relevant. The fusion protein is formed as a result of translocation t(15,17) and is detected in the laboratory by fluorescence in situ hybridization (FISH) or reverse transcriptase polymerase chain reaction (RT-PCR). Diagnostic methods require many laboratory steps with specialized staff. The developed microfluidic assay includes a sandwich enzyme-linked immunosorbent assay (ELISA) system for PML-RARα on surface of magnetic microparticles in a microfluidic chip. A rapid detection of PML-RARα in cell lysates is achieved in less than one hour. A biotinylated PML-antibody on the surface of magnetic streptavidin coated microparticles is used as capture antibody. The bound translocation product is detected by a RARα antibody conjugated with horseradish peroxidase and the substrate QuantaRed. The analysis is performed in microfluidic channels which involves automated liquid processing with stringent washing and short incubation times. The results of the developed assay show that cell lysates of PML-RARα-positive cells (NB-4) can be clearly distinguished from PML-RARα-negative cells (HL-60, MV4-11).</description><subject>Acute myeloid leukemia</subject><subject>Acute promyeloid leukemia</subject><subject>Antibodies</subject><subject>Antigens</subject><subject>Assaying</subject><subject>Automation</subject><subject>Biomarkers, Tumor - genetics</subject><subject>Biomarkers, Tumor - isolation & purification</subject><subject>Chromosomes</subject><subject>Diagnostic systems</subject><subject>Differential diagnosis</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Enzymes</subject><subject>Fluorescence</subject><subject>Fluorescence in situ hybridization</subject><subject>Fusion protein</subject><subject>Genes</subject><subject>Granulocyte Precursor Cells - metabolism</subject><subject>Granulocyte Precursor Cells - pathology</subject><subject>Horseradish peroxidase</subject><subject>Humans</subject><subject>In Situ Hybridization, Fluorescence - methods</subject><subject>Lab-on-a-chip</subject><subject>Laboratories</subject><subject>Leukemia</subject><subject>Leukemia, Promyelocytic, Acute - diagnosis</subject><subject>Leukemia, Promyelocytic, Acute - genetics</subject><subject>Leukemia, Promyelocytic, Acute - pathology</subject><subject>Lysates</subject><subject>Microfluidics</subject><subject>Microfluidics - methods</subject><subject>Oncogene Proteins, Fusion - genetics</subject><subject>Oncogene Proteins, Fusion - isolation & purification</subject><subject>Patients</subject><subject>Peroxidase</subject><subject>Point of care testing</subject><subject>Polymerase chain reaction</subject><subject>Precision Medicine</subject><subject>Promyelocytic Leukemia Protein - genetics</subject><subject>Promyeloid leukemia</subject><subject>Proteins</subject><subject>Retinoic acid</subject><subject>Retinoic Acid Receptor alpha - genetics</subject><subject>RNA-directed DNA polymerase</subject><subject>Streptavidin</subject><subject>Translocation, Genetic - genetics</subject><issn>1422-0067</issn><issn>1661-6596</issn><issn>1422-0067</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>BENPR</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNpVkUtLxDAUhYMovneupeDWah5N0m4E3wojCjrrkKY3Y8a2GZNWnH9vB8dhXJ2Q-3Fybg5CRwSfMVbgczdtIiWU5UVGN9AuyShNMRZyc-28g_ZinGJMGeXFNtphg3Ihsl1UPjkTvK17VzmTXukIVXIDHZjO-TbxNrl8GqWvMzDOOpNcOd_o8AEhJuPo2knSvUNy-62bWQ0L-CX4Zg61N_MOkhH0H9A4fYC2rK4jHC51H43vbt-uH9LR8_3j9eUoNRmhXcoLDNLKnFT5sIwFw5kocWHLgkpRsVJWheUSBKGaVAJnkpR6uCg1FiYvc8720cWv76wvG6gMtF3QtZoFN2SeK6-d-j9p3bua-C8lJc0JLQaDk6VB8J89xE5NfR_aIbOimeBckowvqNNfavi3GAPY1QsEq0Ujar2RAT9eT7WC_ypgP-oniEk</recordid><startdate>20201125</startdate><enddate>20201125</enddate><creator>Emde, Benedikt</creator><creator>Kreher, Heike</creator><creator>Bäumer, Nicole</creator><creator>Bäumer, Sebastian</creator><creator>Bouwes, Dominique</creator><creator>Tickenbrock, Lara</creator><general>MDPI AG</general><general>MDPI</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>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>M2O</scope><scope>MBDVC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>5PM</scope></search><sort><creationdate>20201125</creationdate><title>Microfluidic-Based Detection of AML-Specific Biomarkers Using the Example of Promyelocyte Leukemia</title><author>Emde, Benedikt ; Kreher, Heike ; Bäumer, Nicole ; Bäumer, Sebastian ; Bouwes, Dominique ; Tickenbrock, Lara</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c412t-590e7f781d8238fec536b09fb9276d3b7d9f57e612a1d60471baf57ba06c8b853</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Acute myeloid leukemia</topic><topic>Acute promyeloid leukemia</topic><topic>Antibodies</topic><topic>Antigens</topic><topic>Assaying</topic><topic>Automation</topic><topic>Biomarkers, Tumor - genetics</topic><topic>Biomarkers, Tumor - isolation & purification</topic><topic>Chromosomes</topic><topic>Diagnostic systems</topic><topic>Differential diagnosis</topic><topic>Enzyme-linked immunosorbent assay</topic><topic>Enzymes</topic><topic>Fluorescence</topic><topic>Fluorescence in situ hybridization</topic><topic>Fusion protein</topic><topic>Genes</topic><topic>Granulocyte Precursor Cells - metabolism</topic><topic>Granulocyte Precursor Cells - pathology</topic><topic>Horseradish peroxidase</topic><topic>Humans</topic><topic>In Situ Hybridization, Fluorescence - methods</topic><topic>Lab-on-a-chip</topic><topic>Laboratories</topic><topic>Leukemia</topic><topic>Leukemia, Promyelocytic, Acute - diagnosis</topic><topic>Leukemia, Promyelocytic, Acute - genetics</topic><topic>Leukemia, Promyelocytic, Acute - pathology</topic><topic>Lysates</topic><topic>Microfluidics</topic><topic>Microfluidics - methods</topic><topic>Oncogene Proteins, Fusion - genetics</topic><topic>Oncogene Proteins, Fusion - isolation & purification</topic><topic>Patients</topic><topic>Peroxidase</topic><topic>Point of care testing</topic><topic>Polymerase chain reaction</topic><topic>Precision Medicine</topic><topic>Promyelocytic Leukemia Protein - genetics</topic><topic>Promyeloid leukemia</topic><topic>Proteins</topic><topic>Retinoic acid</topic><topic>Retinoic Acid Receptor alpha - genetics</topic><topic>RNA-directed DNA polymerase</topic><topic>Streptavidin</topic><topic>Translocation, Genetic - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Emde, Benedikt</creatorcontrib><creatorcontrib>Kreher, Heike</creatorcontrib><creatorcontrib>Bäumer, Nicole</creatorcontrib><creatorcontrib>Bäumer, Sebastian</creatorcontrib><creatorcontrib>Bouwes, Dominique</creatorcontrib><creatorcontrib>Tickenbrock, Lara</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>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>ProQuest Central Essentials</collection><collection>ProQuest Central</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>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>Publicly Available Content Database</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>PubMed Central (Full Participant titles)</collection><jtitle>International journal of molecular sciences</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Emde, Benedikt</au><au>Kreher, Heike</au><au>Bäumer, Nicole</au><au>Bäumer, Sebastian</au><au>Bouwes, Dominique</au><au>Tickenbrock, Lara</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Microfluidic-Based Detection of AML-Specific Biomarkers Using the Example of Promyelocyte Leukemia</atitle><jtitle>International journal of molecular sciences</jtitle><addtitle>Int J Mol Sci</addtitle><date>2020-11-25</date><risdate>2020</risdate><volume>21</volume><issue>23</issue><spage>8942</spage><pages>8942-</pages><issn>1422-0067</issn><issn>1661-6596</issn><eissn>1422-0067</eissn><abstract>A microfluidic assay for the detection of promyelocytic leukemia (PML)-retinoic acid receptor α (RARα) fusion protein was developed. This microfluidic-based system can be used for rapid personalized differential diagnosis of acute promyelocyte leukemia (APL) with the aim of early initiation of individualized therapy. The fusion protein PML-RARα occurs in 95% of acute promyelocytic leukemia cases and is considered as diagnostically relevant. The fusion protein is formed as a result of translocation t(15,17) and is detected in the laboratory by fluorescence in situ hybridization (FISH) or reverse transcriptase polymerase chain reaction (RT-PCR). Diagnostic methods require many laboratory steps with specialized staff. The developed microfluidic assay includes a sandwich enzyme-linked immunosorbent assay (ELISA) system for PML-RARα on surface of magnetic microparticles in a microfluidic chip. A rapid detection of PML-RARα in cell lysates is achieved in less than one hour. A biotinylated PML-antibody on the surface of magnetic streptavidin coated microparticles is used as capture antibody. The bound translocation product is detected by a RARα antibody conjugated with horseradish peroxidase and the substrate QuantaRed. The analysis is performed in microfluidic channels which involves automated liquid processing with stringent washing and short incubation times. The results of the developed assay show that cell lysates of PML-RARα-positive cells (NB-4) can be clearly distinguished from PML-RARα-negative cells (HL-60, MV4-11).</abstract><cop>Switzerland</cop><pub>MDPI AG</pub><pmid>33255664</pmid><doi>10.3390/ijms21238942</doi><oa>free_for_read</oa></addata></record> |
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| ispartof | International journal of molecular sciences, 2020-11, Vol.21 (23), p.8942 |
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| source | MDPI - Multidisciplinary Digital Publishing Institute; MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central |
| subjects | Acute myeloid leukemia Acute promyeloid leukemia Antibodies Antigens Assaying Automation Biomarkers, Tumor - genetics Biomarkers, Tumor - isolation & purification Chromosomes Diagnostic systems Differential diagnosis Enzyme-linked immunosorbent assay Enzymes Fluorescence Fluorescence in situ hybridization Fusion protein Genes Granulocyte Precursor Cells - metabolism Granulocyte Precursor Cells - pathology Horseradish peroxidase Humans In Situ Hybridization, Fluorescence - methods Lab-on-a-chip Laboratories Leukemia Leukemia, Promyelocytic, Acute - diagnosis Leukemia, Promyelocytic, Acute - genetics Leukemia, Promyelocytic, Acute - pathology Lysates Microfluidics Microfluidics - methods Oncogene Proteins, Fusion - genetics Oncogene Proteins, Fusion - isolation & purification Patients Peroxidase Point of care testing Polymerase chain reaction Precision Medicine Promyelocytic Leukemia Protein - genetics Promyeloid leukemia Proteins Retinoic acid Retinoic Acid Receptor alpha - genetics RNA-directed DNA polymerase Streptavidin Translocation, Genetic - genetics |
| title | Microfluidic-Based Detection of AML-Specific Biomarkers Using the Example of Promyelocyte Leukemia |
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