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A 3D Microfluidic ELISA for the Detection of Severe Dengue: Sensitivity Improvement and Vroman Effect Amelioration by EDC–NHS Surface Modification

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...

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Autores principales: Maeno, Hinata, Wong, Pooi-Fong, AbuBakar, Sazaly, Yang, Ming, Sam, Sing-Sin, Jamil-Abd, Juraina, Shunmugarajoo, Anusha, Mustafa, Mahiran, Said, Rosaida Md, Mageswaren, Eashwary, Azmel, Azureen, Mat Jelani, Anilawati
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8715748/
https://www.ncbi.nlm.nih.gov/pubmed/34945351
http://dx.doi.org/10.3390/mi12121503
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author Maeno, Hinata
Wong, Pooi-Fong
AbuBakar, Sazaly
Yang, Ming
Sam, Sing-Sin
Jamil-Abd, Juraina
Shunmugarajoo, Anusha
Mustafa, Mahiran
Said, Rosaida Md
Mageswaren, Eashwary
Azmel, Azureen
Mat Jelani, Anilawati
author_facet Maeno, Hinata
Wong, Pooi-Fong
AbuBakar, Sazaly
Yang, Ming
Sam, Sing-Sin
Jamil-Abd, Juraina
Shunmugarajoo, Anusha
Mustafa, Mahiran
Said, Rosaida Md
Mageswaren, Eashwary
Azmel, Azureen
Mat Jelani, Anilawati
author_sort Maeno, Hinata
collection PubMed
description 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—soluble CD163 (sCD163)—within 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–NHS (N-ethyl-N′-(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–NHS coupling prevented antibody desorption and improved the sensitivity. Subsequent comparison of sCD163 detection using a 3D Stack with EDC–NHS coupling and conventional ELISA in dengue patients’ sera revealed a high correlation (R = 0.9298, p < 0.0001) between the two detection platforms. Bland–Altman 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.
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spelling pubmed-87157482021-12-30 A 3D Microfluidic ELISA for the Detection of Severe Dengue: Sensitivity Improvement and Vroman Effect Amelioration by EDC–NHS Surface Modification Maeno, Hinata Wong, Pooi-Fong AbuBakar, Sazaly Yang, Ming Sam, Sing-Sin Jamil-Abd, Juraina Shunmugarajoo, Anusha Mustafa, Mahiran Said, Rosaida Md Mageswaren, Eashwary Azmel, Azureen Mat Jelani, Anilawati Micromachines (Basel) Article 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—soluble CD163 (sCD163)—within 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–NHS (N-ethyl-N′-(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–NHS coupling prevented antibody desorption and improved the sensitivity. Subsequent comparison of sCD163 detection using a 3D Stack with EDC–NHS coupling and conventional ELISA in dengue patients’ sera revealed a high correlation (R = 0.9298, p < 0.0001) between the two detection platforms. Bland–Altman 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. MDPI 2021-11-30 /pmc/articles/PMC8715748/ /pubmed/34945351 http://dx.doi.org/10.3390/mi12121503 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Maeno, Hinata
Wong, Pooi-Fong
AbuBakar, Sazaly
Yang, Ming
Sam, Sing-Sin
Jamil-Abd, Juraina
Shunmugarajoo, Anusha
Mustafa, Mahiran
Said, Rosaida Md
Mageswaren, Eashwary
Azmel, Azureen
Mat Jelani, Anilawati
A 3D Microfluidic ELISA for the Detection of Severe Dengue: Sensitivity Improvement and Vroman Effect Amelioration by EDC–NHS Surface Modification
title A 3D Microfluidic ELISA for the Detection of Severe Dengue: Sensitivity Improvement and Vroman Effect Amelioration by EDC–NHS Surface Modification
title_full A 3D Microfluidic ELISA for the Detection of Severe Dengue: Sensitivity Improvement and Vroman Effect Amelioration by EDC–NHS Surface Modification
title_fullStr A 3D Microfluidic ELISA for the Detection of Severe Dengue: Sensitivity Improvement and Vroman Effect Amelioration by EDC–NHS Surface Modification
title_full_unstemmed A 3D Microfluidic ELISA for the Detection of Severe Dengue: Sensitivity Improvement and Vroman Effect Amelioration by EDC–NHS Surface Modification
title_short A 3D Microfluidic ELISA for the Detection of Severe Dengue: Sensitivity Improvement and Vroman Effect Amelioration by EDC–NHS Surface Modification
title_sort 3d microfluidic elisa for the detection of severe dengue: sensitivity improvement and vroman effect amelioration by edc–nhs surface modification
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8715748/
https://www.ncbi.nlm.nih.gov/pubmed/34945351
http://dx.doi.org/10.3390/mi12121503
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