A bead-based immunogold-silver staining assay on capillary-driven microfluidics

Biomedical Microdevices
Ngoc M PhamEmmanuel Delamarche

Abstract

Point-of-care (POC) diagnostics are critically needed for the detection of infectious diseases, particularly in remote settings where accurate and appropriate diagnosis can save lives. However, it is difficult to implement immunoassays, and specifically immunoassays relying on signal amplification using silver staining, into POC diagnostic devices. Effective immobilization of antibodies in such devices is another challenge. Here, we present strategies for immobilizing capture antibodies (cAbs) in capillary-driven microfluidic chips and implementing a gold-catalyzed silver staining reaction. We illustrate these strategies using a species/anti-species immunoassay and the capillary assembly of fluorescent microbeads functionalized with cAbs in "bead lanes", which are engraved in microfluidic chips. The microfluidic chips are fabricated in silicon (Si) and sealed with a dry film resist. Rabbit IgG antibodies in samples are captured on the beads and bound by detection antibodies (dAbs) conjugated to gold nanoparticles. The gold nanoparticles catalyze the formation of a metallic film of silver, which attenuates fluorescence from the beads in an analyte-concentration dependent manner. The performance of these immunoassays was found co...Continue Reading

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Citations

Jun 8, 2018·The Analyst·Ali Khodayari Bavil, Jungkyu Kim
Nov 23, 2019·Scientific Reports·Marie L SalvaEmmanuel Delamarche
Dec 30, 2020·Biosensors·Pierre-Emmanuel ThirietCarlotta Guiducci
Jul 31, 2021·Journal of Healthcare Engineering·Yuxing ShiWenhao Zhao
Nov 14, 2019·Analytical Chemistry·Jacob B NielsenAdam T Woolley

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