Cargando…

Bead Number Effect in a Magnetic-Beads-Based Digital Microfluidic Immunoassay

In a biomedical diagnosis with a limited sample volume and low concentration, droplet-based microfluidics, also called digital microfluidics, becomes a very attractive approach. Previously, our group developed a magnetic-beads-based digital microfluidic immunoassay with a bead number of around 100,...

Descripción completa

Detalles Bibliográficos
Autores principales: Hsu, Wensyang, Shih, Yu-Teng, Lee, Meng-Shiue, Huang, Hong-Yuan, Wu, Wan-Ning
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9138409/
https://www.ncbi.nlm.nih.gov/pubmed/35624641
http://dx.doi.org/10.3390/bios12050340
_version_ 1784714618296336384
author Hsu, Wensyang
Shih, Yu-Teng
Lee, Meng-Shiue
Huang, Hong-Yuan
Wu, Wan-Ning
author_facet Hsu, Wensyang
Shih, Yu-Teng
Lee, Meng-Shiue
Huang, Hong-Yuan
Wu, Wan-Ning
author_sort Hsu, Wensyang
collection PubMed
description In a biomedical diagnosis with a limited sample volume and low concentration, droplet-based microfluidics, also called digital microfluidics, becomes a very attractive approach. Previously, our group developed a magnetic-beads-based digital microfluidic immunoassay with a bead number of around 100, requiring less than 1 μL of sample volume to achieve a pg/mL level limit of detection (LOD). However, the bead number in each measurement was not the same, causing an unstable coefficient of variation (CV) in the calibration curve. Here, we investigated whether a fixed number of beads in this bead-based digital microfluidic immunoassay could provide more stable results. First, the bead screening chips were developed to extract exactly 100, 49, and 25 magnetic beads with diameters of less than 6 μm. Then, four calibration curves were established. One calibration curve was constructed by using varying bead numbers (50–160) in the process. The other three calibration curves used a fixed number of beads, (100, 49, and 25). The results indicated that the CVs for a fixed number of beads were evidently smaller than the CVs for varying bead numbers, especially in the range of 1 pg/mL to 100 pg/mL, where the CVs for 100 beads were less than 10%. Furthermore, the calculated LOD, based on the composite calibration curves, could be reduced by three orders, from 3.0 pg/mL (for the unfixed bead number) to 0.0287 pg/mL (for 100 beads). However, when the bead numbers were too high (more than 500) or too low (25 or fewer), the bead manipulation for aggregation became more difficult in the magnetic-beads-based digital microfluidic immunoassay chip.
format Online
Article
Text
id pubmed-9138409
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-91384092022-05-28 Bead Number Effect in a Magnetic-Beads-Based Digital Microfluidic Immunoassay Hsu, Wensyang Shih, Yu-Teng Lee, Meng-Shiue Huang, Hong-Yuan Wu, Wan-Ning Biosensors (Basel) Article In a biomedical diagnosis with a limited sample volume and low concentration, droplet-based microfluidics, also called digital microfluidics, becomes a very attractive approach. Previously, our group developed a magnetic-beads-based digital microfluidic immunoassay with a bead number of around 100, requiring less than 1 μL of sample volume to achieve a pg/mL level limit of detection (LOD). However, the bead number in each measurement was not the same, causing an unstable coefficient of variation (CV) in the calibration curve. Here, we investigated whether a fixed number of beads in this bead-based digital microfluidic immunoassay could provide more stable results. First, the bead screening chips were developed to extract exactly 100, 49, and 25 magnetic beads with diameters of less than 6 μm. Then, four calibration curves were established. One calibration curve was constructed by using varying bead numbers (50–160) in the process. The other three calibration curves used a fixed number of beads, (100, 49, and 25). The results indicated that the CVs for a fixed number of beads were evidently smaller than the CVs for varying bead numbers, especially in the range of 1 pg/mL to 100 pg/mL, where the CVs for 100 beads were less than 10%. Furthermore, the calculated LOD, based on the composite calibration curves, could be reduced by three orders, from 3.0 pg/mL (for the unfixed bead number) to 0.0287 pg/mL (for 100 beads). However, when the bead numbers were too high (more than 500) or too low (25 or fewer), the bead manipulation for aggregation became more difficult in the magnetic-beads-based digital microfluidic immunoassay chip. MDPI 2022-05-16 /pmc/articles/PMC9138409/ /pubmed/35624641 http://dx.doi.org/10.3390/bios12050340 Text en © 2022 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
Hsu, Wensyang
Shih, Yu-Teng
Lee, Meng-Shiue
Huang, Hong-Yuan
Wu, Wan-Ning
Bead Number Effect in a Magnetic-Beads-Based Digital Microfluidic Immunoassay
title Bead Number Effect in a Magnetic-Beads-Based Digital Microfluidic Immunoassay
title_full Bead Number Effect in a Magnetic-Beads-Based Digital Microfluidic Immunoassay
title_fullStr Bead Number Effect in a Magnetic-Beads-Based Digital Microfluidic Immunoassay
title_full_unstemmed Bead Number Effect in a Magnetic-Beads-Based Digital Microfluidic Immunoassay
title_short Bead Number Effect in a Magnetic-Beads-Based Digital Microfluidic Immunoassay
title_sort bead number effect in a magnetic-beads-based digital microfluidic immunoassay
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9138409/
https://www.ncbi.nlm.nih.gov/pubmed/35624641
http://dx.doi.org/10.3390/bios12050340
work_keys_str_mv AT hsuwensyang beadnumbereffectinamagneticbeadsbaseddigitalmicrofluidicimmunoassay
AT shihyuteng beadnumbereffectinamagneticbeadsbaseddigitalmicrofluidicimmunoassay
AT leemengshiue beadnumbereffectinamagneticbeadsbaseddigitalmicrofluidicimmunoassay
AT huanghongyuan beadnumbereffectinamagneticbeadsbaseddigitalmicrofluidicimmunoassay
AT wuwanning beadnumbereffectinamagneticbeadsbaseddigitalmicrofluidicimmunoassay