Efficient isolation of encoded microparticles in a degassed micromold for highly sensitive and multiplex immunoassay with signal amplification

Multiplex detection of low-abundance protein biomarkers in biofluids can contribute to diverse biomedical fields such as early diagnosis and precision medicine. However, conventional techniques such as digital ELISA, microarray, and hydrogel-based assay still face limitations in terms of efficient p...

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Bibliographic Details
Published in:Biosensors & bioelectronics Vol. 261; p. 116465
Main Authors: Jang, Wookyoung, Song, E Loomee, Mun, Seok Joon, Bong, Ki Wan
Format: Journal Article
Language:English
Published: England Elsevier B.V 01.10.2024
Subjects:
air
biomarkers
Biomarkers - urine
Biosensing Techniques - methods
biosensors
Chorionic Gonadotropin, beta Subunit, Human - analysis
Chorionic Gonadotropin, beta Subunit, Human - blood
Chorionic Gonadotropin, beta Subunit, Human - isolation & purification
Chorionic Gonadotropin, beta Subunit, Human - urine
detection limit
Dimethylpolysiloxanes - chemistry
early diagnosis
Equipment Design
Female
gonadotropins
Humans
Hydrogel
Immunoassay
Immunoassay - methods
immunoassays
isolation techniques
Limit of Detection
microarray technology
Multiplex detection
nanopores
Particle isolation
peroxidase
phosphates
polydimethylsiloxane
precision medicine
Pregnancy
Protein
proteomics
Signal amplification
urine
Vascular Endothelial Growth Factor A - analysis
Vascular Endothelial Growth Factor A - isolation & purification
Vascular Endothelial Growth Factor A - urine
vascular endothelial growth factors
Immunoassay
Particle isolation
Hydrogel
Multiplex detection
Protein
Signal amplification
ISSN:0956-5663, 1873-4235, 1873-4235
Online Access:Get full text
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Summary:Multiplex detection of low-abundance protein biomarkers in biofluids can contribute to diverse biomedical fields such as early diagnosis and precision medicine. However, conventional techniques such as digital ELISA, microarray, and hydrogel-based assay still face limitations in terms of efficient protein detection due to issues with multiplexing capability, sensitivity, or complicated assay procedures. In this study, we present the degassed micromold-based particle isolation technique for highly sensitive and multiplex immunoassay with enzymatic signal amplification. Using degassing treatment of nanoporous polydimethylsiloxane (PDMS) micromold, the encoded particles are isolated in the mold within 5 min absorbing trapped air bubbles into the mold by air suction capability. Through 10 min of signal amplification in the isolated spaces by fluorogenic substrate and horseradish peroxidase labeled in the particle, the assay signal is amplified with one order of magnitude compared to that of the standard hydrogel-based assay. Using the signal amplification assay, vascular endothelial growth factor (VEGF) and chorionic gonadotropin beta (CG beta), the preeclampsia-related protein biomarkers, are quantitatively detected with a limit of detection (LoD) of 249 fg/mL and 476 fg/mL in phosphate buffer saline. The multiplex immunoassay is conducted to validate negligible non-specific detection signals and robust recovery rates in the multiplex assay. Finally, the VEGF and CG beta in real urine samples are simultaneously and quantitatively detected by the developed assay. Given the high sensitivity, multiplexing capability, and process simplicity, the presented particle isolation-based signal amplification assay holds significant potential in biomedical and proteomic fields.
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ISSN:0956-5663
1873-4235
1873-4235
DOI:10.1016/j.bios.2024.116465