Integrated Single Microbead-Arrayed μ-Fluidic Platform for the Automated Detection of Multiplexed Biomarkers

Abstract

An automated, single microbead-arrayed μ-fluidic immunoassay (AMIA) device is innovatively devised in this study, which enables the highly sensitive and simultaneous detection of multiplex biomarkers with fully automatic operations. The AMIA platform not only achieves automated assay processing and multiplexed target detection by integrating single microbead manipulation, sample loading, multistep washing, and immunoreaction on a microfluidic chip but also confers high sensitivity due to the highly efficient signal enriching effect on a single microbead by the use of only a routine sandwich immunoreaction. As such, as low as the pg/mL level of multiplexed protein biomarkers can be simultaneously determined in a quite small volume of serum (∼20 μL is enough), which can well meet the clinical demand for disease screening and prognosis. What is more, the detection results of several clinically important biomarkers in clinical samples with the AMIA platform exhibit excellent consistency with those obtained by using a standard clinical test. Thus, in virtue of the excellent features in terms of high sensitivity, multiplexing capability, generality, and high degree of automation, the AMIA provides a practical and user-friendly platform for assaying different biomarkers in clinical diagnostics and point-of-care testing.

All on size-coded single bead set: a modular enrich-amplify-amplify strategy for attomolar level multi-immunoassay

“…More fascinatingly, based on a rationally designed size-encoding mechanism (SC-SBEAA), the simultaneous detection of multiple antigens can be accomplished in a single reaction. As a perspective towards real-world clinical applications, we prospect that further integration of the SC-SBEAA with microfluidic chips [33] may provide the most promising and powerful tools for ultrasensitive and multiplexed immunoassay. We envision that if different capture antibody-conjugated single microbeads are packed in a predetermined order/position inside multichannel microfluidic chips, SC-SBEAA can be automatically proceeded for high-throughput target analysis, which will meet the customized needs of basic research or clinical testing in various scenarios and contribute to the human health.…”

Platinum-Decorated Gold Nanoparticle-Based Microfluidic Chip Immunoassay for Ultrasensitive Colorimetric Detection of SARS-CoV-2 Nucleocapsid Protein

“…The microchannel structure ( Figure 1 b) was fabricated in PDMS by rapid prototyping and replication processes as described elsewhere. 30 PDMS membranes and PDMS microchannels were prepared and cut with a certain shape. After the inlet and outlet holes were punched, the PDMS membranes were first bonded with the 3D printed PMMA layers by chemical bonding with a silane coupling agent.…”

“…A height of 100 μm was considered for the chamber. The microchannel structure (Figure b) was fabricated in PDMS by rapid prototyping and replication processes as described elsewhere . PDMS membranes and PDMS microchannels were prepared and cut with a certain shape.…”

The influence of water on size of the monodisperse polystyrene microspheres prepared by dispersion polymerization

“…Micrometer-sized uniform polystyrene microspheres have a wide range of applications in biomedical engineering, biotechnology, electronics, and other fields. [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15] For instance, these precisely sized and monodisperse microparticles can be effectively utilized in tandem with flow cytometry to craft fluorescently encoded microspheres. Moreover, they serve as templates for binding with magnetic nanoparticles, nucleic acid probes, and other biomolecular entities, thus finding application in the field of life sciences.…”