Bead-Based Immunocomplex Entrapment Assays for Rapid, Sensitive, and Multiplexed Detection of Disease Biomarkers with Minimal User Intervention
| Autor: | Yu-Hui Gong, Gerard Marriott |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Immunoassay
Fluid Flow and Transfer Processes Large molecular weight Chemistry Process Chemistry and Technology Bioengineering Bead Multiple target Multiplexing Immunoconjugate visual_art visual_art.visual_art_medium Humans Disease biomarker Disease Instrumentation Biomarkers Diagnostic Techniques and Procedures Biomedical engineering |
| Zdroj: | ACS Sensors. 5:180-190 |
| ISSN: | 2379-3694 |
| Popis: | Current interest in at-home diagnostic devices derives from their potential to disrupt expensive and time-consuming hospital-based diagnostic practices. Conventional immunoassays are often touted for use in at-home diagnostic devices, although in practice they are slow, labor-intensive and require expensive equipment. Here, we introduce bead-based sensors as alternative biomarker detection platforms for at-home diagnostic devices. The immunocomplex entrapment assay (ICEA), and the related enzyme-linked ICEA (ELICEA) offer enhancements over conventional immunoassays in terms of their speed, and minimal requirements for user intervention and instrumentation. In particular, we designed bead-based sensors to entrap large molecular weight complexes between target molecules and signal-generating immunoconjugates while allowing any unbound conjugates to escape from the bead. Confocal fluorescence microscopy was used to demonstrate the sensitivity, robustness, and reproducibility of the ICEA and ELICEA platforms. For example, we showed the intensity of signals generated by entrapped immunoconjugate complexes correlate linearly with the concentration of target molecule in the sample. We employed ICEA, and ELICEA platforms to detect human forms of immunoglobulins, albumin, and κ light chain (KLC). For example, we used ICEA to detect KLC at 5 μg·mL-1 in urine, which would allow for earlier diagnosis of Bence-Jones disease compared to conventional assays. In addition, we showed bead-entrapped phosphatases (AP) in immunocomplexes generate insoluble, blue-colored dyes from AP-substrates that accumulate in beads and allow for visual and cellphone camera-based detection of IgG to 10 ng·mL-1 within 20 min. Finally, we described ICEA and ELICEA platforms to analyze multiple target proteins within individual beads. |
| Databáze: | OpenAIRE |
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