Cargando…
QCM-based assay designs for human serum albumin
Solid-phase synthesis is an elegant way to create molecularly imprinted polymer nanoparticles (nano-MIPs) comprising a single binding site, i.e. mimics of antibodies. When using human serum albumin (HSA) as the template, one achieves nano-MIPs with 53 ± 19 nm diameter, while non-imprinted polymer na...
Autores principales: | , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8748353/ https://www.ncbi.nlm.nih.gov/pubmed/34950982 http://dx.doi.org/10.1007/s00216-021-03771-0 |
_version_ | 1784631002407108608 |
---|---|
author | Sudjarwo, Wisnu Arfian A. Dobler, Mathias Thomas Lieberzeit, Peter A. |
author_facet | Sudjarwo, Wisnu Arfian A. Dobler, Mathias Thomas Lieberzeit, Peter A. |
author_sort | Sudjarwo, Wisnu Arfian A. |
collection | PubMed |
description | Solid-phase synthesis is an elegant way to create molecularly imprinted polymer nanoparticles (nano-MIPs) comprising a single binding site, i.e. mimics of antibodies. When using human serum albumin (HSA) as the template, one achieves nano-MIPs with 53 ± 19 nm diameter, while non-imprinted polymer nanoparticles (nano-NIPs) reach 191 ± 96 nm. Fluorescence assays lead to Stern–Volmer plots revealing selective binding to HSA with selectivity factors of 1.2 compared to bovine serum albumin (BSA), 1.9 for lysozyme, and 4.1 for pepsin. Direct quartz crystal microbalance (QCM) assays confirm these results: nano-MIPs bind to HSA immobilized on QCM surfaces. This opens the way for competitive QCM-based assays for HSA: adding HSA to nanoparticle solutions indeed reduces binding to the QCM surfaces in a concentration-dependent manner. They achieve a limit of detection (LoD) of 80 nM and a limit of quantification (LoQ) of 244 nM. Furthermore, the assay shows recovery rates around 100% for HSA even in the presence of competing analytes. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00216-021-03771-0. |
format | Online Article Text |
id | pubmed-8748353 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-87483532022-01-20 QCM-based assay designs for human serum albumin Sudjarwo, Wisnu Arfian A. Dobler, Mathias Thomas Lieberzeit, Peter A. Anal Bioanal Chem Research Paper Solid-phase synthesis is an elegant way to create molecularly imprinted polymer nanoparticles (nano-MIPs) comprising a single binding site, i.e. mimics of antibodies. When using human serum albumin (HSA) as the template, one achieves nano-MIPs with 53 ± 19 nm diameter, while non-imprinted polymer nanoparticles (nano-NIPs) reach 191 ± 96 nm. Fluorescence assays lead to Stern–Volmer plots revealing selective binding to HSA with selectivity factors of 1.2 compared to bovine serum albumin (BSA), 1.9 for lysozyme, and 4.1 for pepsin. Direct quartz crystal microbalance (QCM) assays confirm these results: nano-MIPs bind to HSA immobilized on QCM surfaces. This opens the way for competitive QCM-based assays for HSA: adding HSA to nanoparticle solutions indeed reduces binding to the QCM surfaces in a concentration-dependent manner. They achieve a limit of detection (LoD) of 80 nM and a limit of quantification (LoQ) of 244 nM. Furthermore, the assay shows recovery rates around 100% for HSA even in the presence of competing analytes. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00216-021-03771-0. Springer Berlin Heidelberg 2021-12-23 2022 /pmc/articles/PMC8748353/ /pubmed/34950982 http://dx.doi.org/10.1007/s00216-021-03771-0 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis 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/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Research Paper Sudjarwo, Wisnu Arfian A. Dobler, Mathias Thomas Lieberzeit, Peter A. QCM-based assay designs for human serum albumin |
title | QCM-based assay designs for human serum albumin |
title_full | QCM-based assay designs for human serum albumin |
title_fullStr | QCM-based assay designs for human serum albumin |
title_full_unstemmed | QCM-based assay designs for human serum albumin |
title_short | QCM-based assay designs for human serum albumin |
title_sort | qcm-based assay designs for human serum albumin |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8748353/ https://www.ncbi.nlm.nih.gov/pubmed/34950982 http://dx.doi.org/10.1007/s00216-021-03771-0 |
work_keys_str_mv | AT sudjarwowisnuarfiana qcmbasedassaydesignsforhumanserumalbumin AT doblermathiasthomas qcmbasedassaydesignsforhumanserumalbumin AT lieberzeitpetera qcmbasedassaydesignsforhumanserumalbumin |