{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T02:35:35Z","timestamp":1760236535678,"version":"build-2065373602"},"reference-count":46,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2021,11,30]],"date-time":"2021-11-30T00:00:00Z","timestamp":1638230400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Energy, Science, Technology, Environment and Climate Change (MESTECC), Malay-sia","award":["FP0514B0025-2 (DSTIN) WorkPackage 5C (GA019-2016)"],"award-info":[{"award-number":["FP0514B0025-2 (DSTIN) WorkPackage 5C (GA019-2016)"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Micromachines"],"abstract":"<jats:p>Serum is commonly used as a specimen in immunoassays but the presence of heterophilic antibodies can potentially interfere with the test results. Previously, we have developed a microfluidic device called: 3D Stack for enzyme-linked immunosorbent assay (ELISA). However, its evaluation was limited to detection from a single protein solution. Here, we investigated the sensitivity of the 3D Stack in detecting a severe dengue biomarker\u2014soluble CD163 (sCD163)\u2014within the serum matrix. To determine potential interactions with serum matrix, a spike-and-recovery assay was performed, using 3D Stacks with and without surface modification by an EDC\u2013NHS (N-ethyl-N\u2032-(3-(dimethylamino)propyl)carbodiimide\/N-hydroxysuccinimide) coupling. Without surface modification, a reduced analyte recovery in proportion to serum concentration was observed because of the Vroman effect, which resulted in competitive displacement of coated capture antibodies by serum proteins with stronger binding affinities. However, EDC\u2013NHS coupling prevented antibody desorption and improved the sensitivity. Subsequent comparison of sCD163 detection using a 3D Stack with EDC\u2013NHS coupling and conventional ELISA in dengue patients\u2019 sera revealed a high correlation (R = 0.9298, p &lt; 0.0001) between the two detection platforms. Bland\u2013Altman analysis further revealed insignificant systematic error between the mean differences of the two methods. These data suggest the potentials of the 3D Stack for further development as a detection platform.<\/jats:p>","DOI":"10.3390\/mi12121503","type":"journal-article","created":{"date-parts":[[2021,11,30]],"date-time":"2021-11-30T22:01:11Z","timestamp":1638309671000},"page":"1503","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["A 3D Microfluidic ELISA for the Detection of Severe Dengue: Sensitivity Improvement and Vroman Effect Amelioration by EDC\u2013NHS Surface Modification"],"prefix":"10.3390","volume":"12","author":[{"given":"Hinata","family":"Maeno","sequence":"first","affiliation":[{"name":"Department of System Design, Tokyo Metropolitan University, Tokyo 191-0065, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6705-2521","authenticated-orcid":false,"given":"Pooi-Fong","family":"Wong","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Faculty of Medicine, University of Malaya, Kuala Lumpur 50603, Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9267-1420","authenticated-orcid":false,"given":"Sazaly","family":"AbuBakar","sequence":"additional","affiliation":[{"name":"Tropical Infectious Diseases Research and Educational Centre (TIDREC), University of Malaya, Kuala Lumpur 50603, Malaysia"},{"name":"WHO Collaborating Centre for Arbovirus Reference and Research (Dengue and Severe Dengue) MAA-12, University of Malaya, Kuala Lumpur 50603, Malaysia"},{"name":"Medical Department, Tengku Ampuan Rahimah Hospital, Klang 41200, Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ming","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of System Design, Tokyo Metropolitan University, Tokyo 191-0065, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2527-5310","authenticated-orcid":false,"given":"Sing-Sin","family":"Sam","sequence":"additional","affiliation":[{"name":"Tropical Infectious Diseases Research and Educational Centre (TIDREC), University of Malaya, Kuala Lumpur 50603, Malaysia"},{"name":"WHO Collaborating Centre for Arbovirus Reference and Research (Dengue and Severe Dengue) MAA-12, University of Malaya, Kuala Lumpur 50603, Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juraina","family":"Jamil-Abd","sequence":"additional","affiliation":[{"name":"Tropical Infectious Diseases Research and Educational Centre (TIDREC), University of Malaya, Kuala Lumpur 50603, Malaysia"},{"name":"WHO Collaborating Centre for Arbovirus Reference and Research (Dengue and Severe Dengue) MAA-12, University of Malaya, Kuala Lumpur 50603, Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anusha","family":"Shunmugarajoo","sequence":"additional","affiliation":[{"name":"Medical Department, Tengku Ampuan Rahimah Hospital, Klang 41200, Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mahiran","family":"Mustafa","sequence":"additional","affiliation":[{"name":"Medical Department, Raja Perempuan Zainab II Hospital, Kota Bharu 15200, Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rosaida Md","family":"Said","sequence":"additional","affiliation":[{"name":"Medical Department, Ampang Hospital, Ampang 68000, Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4747-8541","authenticated-orcid":false,"given":"Eashwary","family":"Mageswaren","sequence":"additional","affiliation":[{"name":"Medical Department, Tengku Ampuan Rahimah Hospital, Klang 41200, Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Azureen","family":"Azmel","sequence":"additional","affiliation":[{"name":"Medical Department, Tengku Ampuan Rahimah Hospital, Klang 41200, Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anilawati","family":"Mat Jelani","sequence":"additional","affiliation":[{"name":"Medical Department, Raja Perempuan Zainab II Hospital, Kota Bharu 15200, Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"871","DOI":"10.1016\/0019-2791(71)90454-X","article-title":"Enzyme-Linked immunosorbent assay (elisa) quantitative assay of immunoglobulin-G","volume":"8","author":"Engvall","year":"1971","journal-title":"Immunochemistry"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1016\/0014-5793(71)80319-8","article-title":"Immunoassay using 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