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Thin Film Plastic Antibody-Based Microplate Assay for Human Serum Albumin Determination
Herein we demonstrate molecularly imprinted polymers (MIP) as plastic antibodies for a microplate-based assay. As the most abundant plasma protein, human serum albumin (HSA) was selected as the target analyte model. Thin film MIP was synthesized by the surface molecular imprinting approach using HSA...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8198403/ https://www.ncbi.nlm.nih.gov/pubmed/34072152 http://dx.doi.org/10.3390/polym13111763 |
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author | Boonsriwong, Worachote Chunta, Suticha Thepsimanon, Nonthawat Singsanan, Sanita Lieberzeit, Peter A. |
author_facet | Boonsriwong, Worachote Chunta, Suticha Thepsimanon, Nonthawat Singsanan, Sanita Lieberzeit, Peter A. |
author_sort | Boonsriwong, Worachote |
collection | PubMed |
description | Herein we demonstrate molecularly imprinted polymers (MIP) as plastic antibodies for a microplate-based assay. As the most abundant plasma protein, human serum albumin (HSA) was selected as the target analyte model. Thin film MIP was synthesized by the surface molecular imprinting approach using HSA as the template. The optimized polymer consisted of acrylic acid (AA) and N-vinylpyrrolidone (VP) in a 2:3 (w/w) ratio, crosslinked with N,N′-(1,2-dihydroxyethylene) bisacrylamide (DHEBA) and then coated on the microplate well. The binding of MIP toward the bound HSA was achieved via the Bradford reaction. The assay revealed a dynamic detection range toward HSA standards in the clinically relevant 1–10 g/dL range, with a 0.01 g/dL detection limit. HSA-MIP showed minimal interference from other serum protein components: γ-globulin had 11% of the HSA response, α-globulin of high-density lipoprotein had 9%, and β-globulin of low-density lipoprotein had 7%. The analytical accuracy of the assay was 89–106% at the 95% confidence interval, with precision at 4–9%. The MIP-coated microplate was stored for 2 months at room temperature without losing its binding ability. The results suggest that the thin film plastic antibody system can be successfully applied to analytical/pseudoimmunological HSA determinations in clinical applications. |
format | Online Article Text |
id | pubmed-8198403 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-81984032021-06-14 Thin Film Plastic Antibody-Based Microplate Assay for Human Serum Albumin Determination Boonsriwong, Worachote Chunta, Suticha Thepsimanon, Nonthawat Singsanan, Sanita Lieberzeit, Peter A. Polymers (Basel) Article Herein we demonstrate molecularly imprinted polymers (MIP) as plastic antibodies for a microplate-based assay. As the most abundant plasma protein, human serum albumin (HSA) was selected as the target analyte model. Thin film MIP was synthesized by the surface molecular imprinting approach using HSA as the template. The optimized polymer consisted of acrylic acid (AA) and N-vinylpyrrolidone (VP) in a 2:3 (w/w) ratio, crosslinked with N,N′-(1,2-dihydroxyethylene) bisacrylamide (DHEBA) and then coated on the microplate well. The binding of MIP toward the bound HSA was achieved via the Bradford reaction. The assay revealed a dynamic detection range toward HSA standards in the clinically relevant 1–10 g/dL range, with a 0.01 g/dL detection limit. HSA-MIP showed minimal interference from other serum protein components: γ-globulin had 11% of the HSA response, α-globulin of high-density lipoprotein had 9%, and β-globulin of low-density lipoprotein had 7%. The analytical accuracy of the assay was 89–106% at the 95% confidence interval, with precision at 4–9%. The MIP-coated microplate was stored for 2 months at room temperature without losing its binding ability. The results suggest that the thin film plastic antibody system can be successfully applied to analytical/pseudoimmunological HSA determinations in clinical applications. MDPI 2021-05-27 /pmc/articles/PMC8198403/ /pubmed/34072152 http://dx.doi.org/10.3390/polym13111763 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 Boonsriwong, Worachote Chunta, Suticha Thepsimanon, Nonthawat Singsanan, Sanita Lieberzeit, Peter A. Thin Film Plastic Antibody-Based Microplate Assay for Human Serum Albumin Determination |
title | Thin Film Plastic Antibody-Based Microplate Assay for Human Serum Albumin Determination |
title_full | Thin Film Plastic Antibody-Based Microplate Assay for Human Serum Albumin Determination |
title_fullStr | Thin Film Plastic Antibody-Based Microplate Assay for Human Serum Albumin Determination |
title_full_unstemmed | Thin Film Plastic Antibody-Based Microplate Assay for Human Serum Albumin Determination |
title_short | Thin Film Plastic Antibody-Based Microplate Assay for Human Serum Albumin Determination |
title_sort | thin film plastic antibody-based microplate assay for human serum albumin determination |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8198403/ https://www.ncbi.nlm.nih.gov/pubmed/34072152 http://dx.doi.org/10.3390/polym13111763 |
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