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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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Main Authors:	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
Format:	Article
Published:	MDPI 2021
Subjects:	Q Science (General) QD Chemistry
Online Access:	http://eprints.um.edu.my/34535/
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spelling	my.um.eprints.345352022-06-01T02:59:01Z http://eprints.um.edu.my/34535/ 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 Q Science (General) QD Chemistry 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 2021-12 Article PeerReviewed Maeno, Hinata and Wong, Pooi-Fong and AbuBakar, Sazaly and Yang, Ming and Sam, Sing-Sin and Jamil-Abd, Juraina and Shunmugarajoo, Anusha and Mustafa, Mahiran and Said, Rosaida Md and Mageswaren, Eashwary and Azmel, Azureen and Mat Jelani, Anilawati (2021) A 3D microfluidic elisa for the detection of severe dengue: sensitivity improvement and vroman effect amelioration by edc-nhs surface modification. Micromachines, 12 (12). ISSN 2072-666X, DOI https://doi.org/10.3390/mi12121503 <https://doi.org/10.3390/mi12121503>. 10.3390/mi12121503
institution	Universiti Malaya
building	UM Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Malaya
content_source	UM Research Repository
url_provider	http://eprints.um.edu.my/
topic	Q Science (General) QD Chemistry
spellingShingle	Q Science (General) QD Chemistry 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
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.
format	Article
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
title	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_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_sort	3d microfluidic elisa for the detection of severe dengue: sensitivity improvement and vroman effect amelioration by edc-nhs surface modification
publisher	MDPI
publishDate	2021
url	http://eprints.um.edu.my/34535/
_version_	1735409612434178048
score	13.211869

A 3D microfluidic elisa for the detection of severe dengue: sensitivity improvement and vroman effect amelioration by edc-nhs surface modification

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