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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Vydáno v:	Biosensors & bioelectronics Ročník 261; s. 116465
Hlavní autoři:	Jang, Wookyoung, Song, E Loomee, Mun, Seok Joon, Bong, Ki Wan
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	England Elsevier B.V 01.10.2024
Témata:	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
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Shrnutí:	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.
Bibliografie:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0956-5663 1873-4235 1873-4235
DOI:	10.1016/j.bios.2024.116465