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

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

Biosens Bioelectron

Department of Chemical and Biological Engineering, Korea University, Seoul, 02841, Republic of Korea. Electronic address:

Published: October 2024

AI Article Synopsis

  • The study introduces a new technique for detecting low-abundance protein biomarkers in biofluids, which is important for early diagnosis and precision medicine.
  • The method, called degassed micromold-based particle isolation, enhances sensitivity and multiplexing capability by isolating particles in a PDMS mold and using signal amplification with an enzyme.
  • This new approach demonstrated significant improvements in detection limits for specific biomarkers related to preeclampsia, showing potential applications in various biomedical and proteomic fields.

Article Abstract

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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Source
http://dx.doi.org/10.1016/j.bios.2024.116465DOI Listing

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