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Point-of-care vertical flow allergen microarray assay: proof of concept
Sophisticated equipment, lengthy protocols, and skilled operators are required to perform protein microarray-based affinity assays. Consequently, novel tools are needed to bring biomarkers and biomarker panels into clinical use in different settings. Here, we describe a novel paper-based vertical fl...
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Published in: | Clinical chemistry (Baltimore, Md.) Md.), 2014-09, Vol.60 (9), p.1209-1216 |
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creator | Chinnasamy, Thiruppathiraja Segerink, Loes I Nystrand, Mats Gantelius, Jesper Andersson Svahn, Helene |
description | Sophisticated equipment, lengthy protocols, and skilled operators are required to perform protein microarray-based affinity assays. Consequently, novel tools are needed to bring biomarkers and biomarker panels into clinical use in different settings. Here, we describe a novel paper-based vertical flow microarray (VFM) system with a multiplexing capacity of at least 1480 microspot binding sites, colorimetric readout, high sensitivity, and assay time of |
doi_str_mv | 10.1373/clinchem.2014.223230 |
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Affinity binders were deposited on nitrocellulose membranes by conventional microarray printing. Buffers and reagents were applied vertically by use of a flow controlled syringe pump. As a clinical model system, we analyzed 31 precharacterized human serum samples using the array system with 10 allergen components to detect specific IgE reactivities. We detected bound analytes using gold nanoparticle conjugates with assay time of ≤10 min. Microarray images were captured by a consumer-grade flatbed scanner.
A sensitivity of 1 ng/mL was demonstrated with the VFM assay with colorimetric readout. The reproducibility (CV) of the system was <14%. The observed concordance with a clinical assay, ImmunoCAP, was R(2) = 0.89 (n = 31).
In this proof-of-concept study, we demonstrated that the VFM assay, which combines features from protein microarrays and paper-based colorimetric systems, could offer an interesting alternative for future highly multiplexed affinity point-of-care testing.</description><identifier>ISSN: 0009-9147</identifier><identifier>ISSN: 1530-8561</identifier><identifier>EISSN: 1530-8561</identifier><identifier>DOI: 10.1373/clinchem.2014.223230</identifier><identifier>PMID: 25006224</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>Accuracy ; Allergens ; Allergens - analysis ; Allergies ; Binders ; Colorimetry ; Data processing ; Flow control ; Gold ; Gold - chemistry ; Humans ; Immunoassay ; Immunoglobulin E ; Immunoglobulin E - blood ; Laboratories ; Membranes ; Metal Nanoparticles ; Metal Nanoparticles - chemistry ; Nanoparticles ; Point-of-Care Systems ; Protein Array Analysis ; Proteins ; Reagents ; Reproducibility of Results ; Sensitivity and Specificity ; Vertical flow</subject><ispartof>Clinical chemistry (Baltimore, Md.), 2014-09, Vol.60 (9), p.1209-1216</ispartof><rights>2014 American Association for Clinical Chemistry.</rights><rights>Copyright American Association for Clinical Chemistry Sep 2014</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c485t-2e0f619ff8d8849d1e2b47b97229a0b293dfb555259a21d9d1eb57327b077cc43</citedby><cites>FETCH-LOGICAL-c485t-2e0f619ff8d8849d1e2b47b97229a0b293dfb555259a21d9d1eb57327b077cc43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,776,780,881,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25006224$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-157573$$DView record from Swedish Publication Index$$Hfree_for_read</backlink></links><search><creatorcontrib>Chinnasamy, Thiruppathiraja</creatorcontrib><creatorcontrib>Segerink, Loes I</creatorcontrib><creatorcontrib>Nystrand, Mats</creatorcontrib><creatorcontrib>Gantelius, Jesper</creatorcontrib><creatorcontrib>Andersson Svahn, Helene</creatorcontrib><title>Point-of-care vertical flow allergen microarray assay: proof of concept</title><title>Clinical chemistry (Baltimore, Md.)</title><addtitle>Clin Chem</addtitle><description>Sophisticated equipment, lengthy protocols, and skilled operators are required to perform protein microarray-based affinity assays. Consequently, novel tools are needed to bring biomarkers and biomarker panels into clinical use in different settings. Here, we describe a novel paper-based vertical flow microarray (VFM) system with a multiplexing capacity of at least 1480 microspot binding sites, colorimetric readout, high sensitivity, and assay time of <10 min before imaging and data analysis.
Affinity binders were deposited on nitrocellulose membranes by conventional microarray printing. Buffers and reagents were applied vertically by use of a flow controlled syringe pump. As a clinical model system, we analyzed 31 precharacterized human serum samples using the array system with 10 allergen components to detect specific IgE reactivities. We detected bound analytes using gold nanoparticle conjugates with assay time of ≤10 min. Microarray images were captured by a consumer-grade flatbed scanner.
A sensitivity of 1 ng/mL was demonstrated with the VFM assay with colorimetric readout. The reproducibility (CV) of the system was <14%. The observed concordance with a clinical assay, ImmunoCAP, was R(2) = 0.89 (n = 31).
In this proof-of-concept study, we demonstrated that the VFM assay, which combines features from protein microarrays and paper-based colorimetric systems, could offer an interesting alternative for future highly multiplexed affinity point-of-care testing.</description><subject>Accuracy</subject><subject>Allergens</subject><subject>Allergens - analysis</subject><subject>Allergies</subject><subject>Binders</subject><subject>Colorimetry</subject><subject>Data processing</subject><subject>Flow control</subject><subject>Gold</subject><subject>Gold - chemistry</subject><subject>Humans</subject><subject>Immunoassay</subject><subject>Immunoglobulin E</subject><subject>Immunoglobulin E - blood</subject><subject>Laboratories</subject><subject>Membranes</subject><subject>Metal Nanoparticles</subject><subject>Metal Nanoparticles - chemistry</subject><subject>Nanoparticles</subject><subject>Point-of-Care Systems</subject><subject>Protein Array Analysis</subject><subject>Proteins</subject><subject>Reagents</subject><subject>Reproducibility of Results</subject><subject>Sensitivity and Specificity</subject><subject>Vertical flow</subject><issn>0009-9147</issn><issn>1530-8561</issn><issn>1530-8561</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><recordid>eNpdkU9LwzAYh4Mobk6_gUjBi5fO_G1ab2PqFAZ6UK8hTZOts21m0jr27c3otoMQCIHn9_Lm9wBwjeAYEU7uVVU2aqnrMYaIjjEmmMATMESMwDhlCToFQwhhFmeI8gG48H4VnpSnyTkYYAZhgjEdgtm7LZs2tiZW0unoV7u2VLKKTGU3kawq7Ra6iepSOSudk9tIei-3D9HaWWuicJRtlF63l-DMyMrrq_09Ap_PTx_Tl3j-NnudTuaxoilrY6yhSVBmTFqkKc0KpHFOeZ5xjDMJc5yRwuSMMcwyiVGxA3LGCeY55FwpSkYg7uf6jV53uVi7spZuK6wsxWP5NRHWLcR3uxSI8RAM_F3Ph4V_Ou1bUZde6aqSjbadDxhLGeIUwoDe_kNXtnNN-I1ACaKhOxJaHgHaU6ER7502xxUQFDsx4iBG7MSIXkyI3eyHd3mti2PoYIL8AWiYigo</recordid><startdate>20140901</startdate><enddate>20140901</enddate><creator>Chinnasamy, Thiruppathiraja</creator><creator>Segerink, Loes I</creator><creator>Nystrand, Mats</creator><creator>Gantelius, Jesper</creator><creator>Andersson Svahn, Helene</creator><general>Oxford University Press</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>4U-</scope><scope>7QO</scope><scope>7RV</scope><scope>7TM</scope><scope>7U7</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>BKSAR</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>NAPCQ</scope><scope>P64</scope><scope>PCBAR</scope><scope>PDBOC</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>S0X</scope><scope>7X8</scope><scope>ADTPV</scope><scope>AOWAS</scope><scope>D8V</scope></search><sort><creationdate>20140901</creationdate><title>Point-of-care vertical flow allergen microarray assay: proof of concept</title><author>Chinnasamy, Thiruppathiraja ; Segerink, Loes I ; Nystrand, Mats ; Gantelius, Jesper ; Andersson Svahn, Helene</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c485t-2e0f619ff8d8849d1e2b47b97229a0b293dfb555259a21d9d1eb57327b077cc43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Accuracy</topic><topic>Allergens</topic><topic>Allergens - 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Academic</collection><collection>SwePub</collection><collection>SwePub Articles</collection><collection>SWEPUB Kungliga Tekniska Högskolan</collection><jtitle>Clinical chemistry (Baltimore, Md.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chinnasamy, Thiruppathiraja</au><au>Segerink, Loes I</au><au>Nystrand, Mats</au><au>Gantelius, Jesper</au><au>Andersson Svahn, Helene</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Point-of-care vertical flow allergen microarray assay: proof of concept</atitle><jtitle>Clinical chemistry (Baltimore, Md.)</jtitle><addtitle>Clin Chem</addtitle><date>2014-09-01</date><risdate>2014</risdate><volume>60</volume><issue>9</issue><spage>1209</spage><epage>1216</epage><pages>1209-1216</pages><issn>0009-9147</issn><issn>1530-8561</issn><eissn>1530-8561</eissn><abstract>Sophisticated equipment, lengthy protocols, and skilled operators are required to perform protein microarray-based affinity assays. Consequently, novel tools are needed to bring biomarkers and biomarker panels into clinical use in different settings. Here, we describe a novel paper-based vertical flow microarray (VFM) system with a multiplexing capacity of at least 1480 microspot binding sites, colorimetric readout, high sensitivity, and assay time of <10 min before imaging and data analysis.
Affinity binders were deposited on nitrocellulose membranes by conventional microarray printing. Buffers and reagents were applied vertically by use of a flow controlled syringe pump. As a clinical model system, we analyzed 31 precharacterized human serum samples using the array system with 10 allergen components to detect specific IgE reactivities. We detected bound analytes using gold nanoparticle conjugates with assay time of ≤10 min. Microarray images were captured by a consumer-grade flatbed scanner.
A sensitivity of 1 ng/mL was demonstrated with the VFM assay with colorimetric readout. The reproducibility (CV) of the system was <14%. The observed concordance with a clinical assay, ImmunoCAP, was R(2) = 0.89 (n = 31).
In this proof-of-concept study, we demonstrated that the VFM assay, which combines features from protein microarrays and paper-based colorimetric systems, could offer an interesting alternative for future highly multiplexed affinity point-of-care testing.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>25006224</pmid><doi>10.1373/clinchem.2014.223230</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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source | Oxford Journals Online |
subjects | Accuracy Allergens Allergens - analysis Allergies Binders Colorimetry Data processing Flow control Gold Gold - chemistry Humans Immunoassay Immunoglobulin E Immunoglobulin E - blood Laboratories Membranes Metal Nanoparticles Metal Nanoparticles - chemistry Nanoparticles Point-of-Care Systems Protein Array Analysis Proteins Reagents Reproducibility of Results Sensitivity and Specificity Vertical flow |
title | Point-of-care vertical flow allergen microarray assay: proof of concept |
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