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Handheld pH meter–assisted immunoassay for C-reactive protein using glucose oxidase–conjugated dendrimer loaded with platinum nanozymes

A simple and feasible pH meter–based immunoassay is reported for detection of C-reactive protein (CRP) using glucose oxidase (GOD)–conjugated dendrimer loaded with platinum nanozyme. Initially, platinum nanozymes were loaded into the dendrimers through an in situ synthetic method. Then, GOD and mono...

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Autores principales: Li, Bin, Ge, Lilin, Lyu, Peng, Chen, Meijuan, Zhang, Xiongfei, Xie, Shuping, Wu, Qinan, Kwok, Hang Fai
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Vienna 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7779416/
https://www.ncbi.nlm.nih.gov/pubmed/33389237
http://dx.doi.org/10.1007/s00604-020-04687-9
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author Li, Bin
Ge, Lilin
Lyu, Peng
Chen, Meijuan
Zhang, Xiongfei
Xie, Shuping
Wu, Qinan
Kwok, Hang Fai
author_facet Li, Bin
Ge, Lilin
Lyu, Peng
Chen, Meijuan
Zhang, Xiongfei
Xie, Shuping
Wu, Qinan
Kwok, Hang Fai
author_sort Li, Bin
collection PubMed
description A simple and feasible pH meter–based immunoassay is reported for detection of C-reactive protein (CRP) using glucose oxidase (GOD)–conjugated dendrimer loaded with platinum nanozyme. Initially, platinum nanozymes were loaded into the dendrimers through an in situ synthetic method. Then, GOD and monoclonal anti-CRP antibody with a high molar ratio were covalently conjugated onto carboxylated dendrimers via typical carbodiimide coupling. The immunoreaction was carried out with a competitive mode in a CRP-coated microplate. Along with formation of immunocomplex, the added glucose was oxidized into gluconic acid and hydrogen peroxide by GOD, and the latter was further decomposed by platinum nanozyme, thus accelerating chemical reaction in the positive direction. The produced gluconic acid changed the pH of detection solution, which was determined using a handheld pH meter. Under optimum conditions, the pH meter–based immunoassay gave a good signal toward target CRP from 0.01 to 100 ng mL(−1). The limit of detection was 5.9 pg mL(−1). An intermediate precision ≤ 11.2% was acquired with batch-to-batch identification. No nonspecific adsorption was observed during a series of procedures to detect target CRP, and the cross-reaction against other biomarkers was very low. Importantly, our system gave well-matched results for analysis of human serum samples relative to a referenced ELISA kit. Graphical abstract [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00604-020-04687-9.
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spelling pubmed-77794162021-01-11 Handheld pH meter–assisted immunoassay for C-reactive protein using glucose oxidase–conjugated dendrimer loaded with platinum nanozymes Li, Bin Ge, Lilin Lyu, Peng Chen, Meijuan Zhang, Xiongfei Xie, Shuping Wu, Qinan Kwok, Hang Fai Mikrochim Acta Original Paper A simple and feasible pH meter–based immunoassay is reported for detection of C-reactive protein (CRP) using glucose oxidase (GOD)–conjugated dendrimer loaded with platinum nanozyme. Initially, platinum nanozymes were loaded into the dendrimers through an in situ synthetic method. Then, GOD and monoclonal anti-CRP antibody with a high molar ratio were covalently conjugated onto carboxylated dendrimers via typical carbodiimide coupling. The immunoreaction was carried out with a competitive mode in a CRP-coated microplate. Along with formation of immunocomplex, the added glucose was oxidized into gluconic acid and hydrogen peroxide by GOD, and the latter was further decomposed by platinum nanozyme, thus accelerating chemical reaction in the positive direction. The produced gluconic acid changed the pH of detection solution, which was determined using a handheld pH meter. Under optimum conditions, the pH meter–based immunoassay gave a good signal toward target CRP from 0.01 to 100 ng mL(−1). The limit of detection was 5.9 pg mL(−1). An intermediate precision ≤ 11.2% was acquired with batch-to-batch identification. No nonspecific adsorption was observed during a series of procedures to detect target CRP, and the cross-reaction against other biomarkers was very low. Importantly, our system gave well-matched results for analysis of human serum samples relative to a referenced ELISA kit. Graphical abstract [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00604-020-04687-9. Springer Vienna 2021-01-03 2021 /pmc/articles/PMC7779416/ /pubmed/33389237 http://dx.doi.org/10.1007/s00604-020-04687-9 Text en © The Author(s) 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Original Paper
Li, Bin
Ge, Lilin
Lyu, Peng
Chen, Meijuan
Zhang, Xiongfei
Xie, Shuping
Wu, Qinan
Kwok, Hang Fai
Handheld pH meter–assisted immunoassay for C-reactive protein using glucose oxidase–conjugated dendrimer loaded with platinum nanozymes
title Handheld pH meter–assisted immunoassay for C-reactive protein using glucose oxidase–conjugated dendrimer loaded with platinum nanozymes
title_full Handheld pH meter–assisted immunoassay for C-reactive protein using glucose oxidase–conjugated dendrimer loaded with platinum nanozymes
title_fullStr Handheld pH meter–assisted immunoassay for C-reactive protein using glucose oxidase–conjugated dendrimer loaded with platinum nanozymes
title_full_unstemmed Handheld pH meter–assisted immunoassay for C-reactive protein using glucose oxidase–conjugated dendrimer loaded with platinum nanozymes
title_short Handheld pH meter–assisted immunoassay for C-reactive protein using glucose oxidase–conjugated dendrimer loaded with platinum nanozymes
title_sort handheld ph meter–assisted immunoassay for c-reactive protein using glucose oxidase–conjugated dendrimer loaded with platinum nanozymes
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7779416/
https://www.ncbi.nlm.nih.gov/pubmed/33389237
http://dx.doi.org/10.1007/s00604-020-04687-9
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