Development of a Novel Homogeneous Nanoparticle-Based Assay for Rapid and High-Throughput Quantitation of the sST2 Protein in Human Serum

Shenxia Gao, Junpu Li The Clinical Laboratory, Tianjin Chest Hospital, Tianjin, People’s Republic of ChinaCorrespondence: Junpu LiThe Clinical Laboratory, Tianjin Chest Hospital, No. 261 Taierzhuang South Road, Jinnan District, Tianjin 300222, People’s Republic of ChinaTel/Fax +8...

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Autores principales:	Gao S, Li J
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2020
Materias:	sst2 nanoparticles homogeneous chemiluminescence immunoassay Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/281094471a5744ecb0dbb9c648037d57
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spelling	oai:doaj.org-article:281094471a5744ecb0dbb9c648037d572021-12-02T16:01:14ZDevelopment of a Novel Homogeneous Nanoparticle-Based Assay for Rapid and High-Throughput Quantitation of the sST2 Protein in Human Serum1178-2013https://doaj.org/article/281094471a5744ecb0dbb9c648037d572020-12-01T00:00:00Zhttps://www.dovepress.com/development-of-a-novel-homogeneous-nanoparticle-based-assay-for-rapid--peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Shenxia Gao, Junpu Li The Clinical Laboratory, Tianjin Chest Hospital, Tianjin, People’s Republic of ChinaCorrespondence: Junpu LiThe Clinical Laboratory, Tianjin Chest Hospital, No. 261 Taierzhuang South Road, Jinnan District, Tianjin 300222, People’s Republic of ChinaTel/Fax +86 22-88185111Email junpuli@163.comPurpose: The suppression of tumorigenicity 2 (ST2) protein is a member of the interleukin-1 receptor family with the transmembrane (ST2L) and soluble (sST2) subtypes and plays an important role in several diseases. Therefore, the present study aimed to establish and validate a novel amplified luminescent proximity homogeneous immunoassay (AlphaLISA) for the detection of sST2 in human serum.Materials and Methods: Based on a sandwich-type immunoassay format, sST2 was captured using two different anti-sST2 antibodies. One of the antibodies was biotinylated while the other one was coated with AlphaLISA chemibeads. Thereafter, multiple tests were conducted to optimize the working conditions and validate analytical performance.Results: The optimum concentration of the biotinylated antibodies was 1 μg/mL while the optimal dilution ratio for the anti-sST2 antibodies and conjugated chemibeads was 1:500. In addition, the optimal antigen-antibody reaction time was 15 minutes (min). Notably, the developed method showed a short turnaround time of about 25 min. Moreover, the assay exhibited high sensitivity with a limit of detection (LOD) of 0.176 ng/mL and a limit of quantification (LOQ) of 0.8 ng/mL. Furthermore, the intra-assay precision and inter-assay precision values were 5.29– 7.10% and 9.41%– 13.66%, respectively. It is also noteworthy that the test results deviated by less than ± 10% when samples had ≤ 10.0 ng/mL of triglycerides, ≤ 0.5 mmol/L of bilirubin, ≤ 5.0 g/L of triglyceride, and ≤ 250 μg/L of biotin. Additionally, the developed assay was almost consistent with the commercially available PresageTM ST2 assay kit, with a Spearman correlation coefficient of 0.916 and an R2 of 0.963 as well as a slope of 0.957 from linear regression analysis.Conclusion: The present study showed that the sandwich AlphaLISA is a rapid, high-throughput, and reliable test for studying the levels of sST2 in a variety of diseases.Keywords: sST2, nanoparticles, homogeneous, chemiluminescence immunoassayGao SLi JDove Medical Pressarticlesst2nanoparticleshomogeneouschemiluminescence immunoassayMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 15, Pp 10539-10546 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	sst2 nanoparticles homogeneous chemiluminescence immunoassay Medicine (General) R5-920
spellingShingle	sst2 nanoparticles homogeneous chemiluminescence immunoassay Medicine (General) R5-920 Gao S Li J Development of a Novel Homogeneous Nanoparticle-Based Assay for Rapid and High-Throughput Quantitation of the sST2 Protein in Human Serum
description	Shenxia Gao, Junpu Li The Clinical Laboratory, Tianjin Chest Hospital, Tianjin, People’s Republic of ChinaCorrespondence: Junpu LiThe Clinical Laboratory, Tianjin Chest Hospital, No. 261 Taierzhuang South Road, Jinnan District, Tianjin 300222, People’s Republic of ChinaTel/Fax +86 22-88185111Email junpuli@163.comPurpose: The suppression of tumorigenicity 2 (ST2) protein is a member of the interleukin-1 receptor family with the transmembrane (ST2L) and soluble (sST2) subtypes and plays an important role in several diseases. Therefore, the present study aimed to establish and validate a novel amplified luminescent proximity homogeneous immunoassay (AlphaLISA) for the detection of sST2 in human serum.Materials and Methods: Based on a sandwich-type immunoassay format, sST2 was captured using two different anti-sST2 antibodies. One of the antibodies was biotinylated while the other one was coated with AlphaLISA chemibeads. Thereafter, multiple tests were conducted to optimize the working conditions and validate analytical performance.Results: The optimum concentration of the biotinylated antibodies was 1 μg/mL while the optimal dilution ratio for the anti-sST2 antibodies and conjugated chemibeads was 1:500. In addition, the optimal antigen-antibody reaction time was 15 minutes (min). Notably, the developed method showed a short turnaround time of about 25 min. Moreover, the assay exhibited high sensitivity with a limit of detection (LOD) of 0.176 ng/mL and a limit of quantification (LOQ) of 0.8 ng/mL. Furthermore, the intra-assay precision and inter-assay precision values were 5.29– 7.10% and 9.41%– 13.66%, respectively. It is also noteworthy that the test results deviated by less than ± 10% when samples had ≤ 10.0 ng/mL of triglycerides, ≤ 0.5 mmol/L of bilirubin, ≤ 5.0 g/L of triglyceride, and ≤ 250 μg/L of biotin. Additionally, the developed assay was almost consistent with the commercially available PresageTM ST2 assay kit, with a Spearman correlation coefficient of 0.916 and an R2 of 0.963 as well as a slope of 0.957 from linear regression analysis.Conclusion: The present study showed that the sandwich AlphaLISA is a rapid, high-throughput, and reliable test for studying the levels of sST2 in a variety of diseases.Keywords: sST2, nanoparticles, homogeneous, chemiluminescence immunoassay
format	article
author	Gao S Li J
author_facet	Gao S Li J
author_sort	Gao S
title	Development of a Novel Homogeneous Nanoparticle-Based Assay for Rapid and High-Throughput Quantitation of the sST2 Protein in Human Serum
title_short	Development of a Novel Homogeneous Nanoparticle-Based Assay for Rapid and High-Throughput Quantitation of the sST2 Protein in Human Serum
title_full	Development of a Novel Homogeneous Nanoparticle-Based Assay for Rapid and High-Throughput Quantitation of the sST2 Protein in Human Serum
title_fullStr	Development of a Novel Homogeneous Nanoparticle-Based Assay for Rapid and High-Throughput Quantitation of the sST2 Protein in Human Serum
title_full_unstemmed	Development of a Novel Homogeneous Nanoparticle-Based Assay for Rapid and High-Throughput Quantitation of the sST2 Protein in Human Serum
title_sort	development of a novel homogeneous nanoparticle-based assay for rapid and high-throughput quantitation of the sst2 protein in human serum
publisher	Dove Medical Press
publishDate	2020
url	https://doaj.org/article/281094471a5744ecb0dbb9c648037d57
work_keys_str_mv	AT gaos developmentofanovelhomogeneousnanoparticlebasedassayforrapidandhighthroughputquantitationofthesst2proteininhumanserum AT lij developmentofanovelhomogeneousnanoparticlebasedassayforrapidandhighthroughputquantitationofthesst2proteininhumanserum
_version_	1718385260261015552

Development of a Novel Homogeneous Nanoparticle-Based Assay for Rapid and High-Throughput Quantitation of the sST2 Protein in Human Serum

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