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Published on: June 28, 2024
Functionalized fluorescent core-shell nanoparticles used as a fluorescent labels in fluoroimmunoassay for IL-6
1Department of Chemistry, School of Chemistry and Materials Science, Shaanxi Normal University, Xi'an 710062, China.
Insights
A new fluoroimmunoassay uses functionalized fluorescent nanoparticles to detect interleukin-6 (IL-6). This sensitive method accurately quantifies IL-6 in samples, offering a simple and reproducible diagnostic tool.
Area of Science:
- Biomolecular assays
- Nanotechnology in diagnostics
- Fluorescence-based detection
Background:
- Biomolecule-conjugated nanoparticle labels are crucial for various assay applications.
- Accurate detection of biomarkers like interleukin-6 (IL-6) is vital in diagnostics.
Purpose of the Study:
- To develop a sensitive fluoroimmunoassay for recombinant human interleukin-6 (IL-6) detection.
- To utilize functionalized Rubpy-encapsulated fluorescent core-shell silica nanoparticles for IL-6 labeling.
Main Methods:
- A sandwich fluoroimmunoassay format was employed, capturing IL-6 antigen.
- Functionalized fluorescent core-shell silica nanoparticles conjugated with anti-IL-6 monoclonal antibodies were used as labels.
- Fluorescence microscopy imaging was explored for antibody labeling and assay visualization.
Main Results:
- The assay demonstrated a linear calibration range of 20-1250 pg/mL for IL-6.
- A low detection limit of 7 pg/mL (3 sigma) was achieved.
- High reproducibility was confirmed with a relative standard deviation of 3.2% for IL-6 measurements.
Conclusions:
- The proposed method offers high sensitivity, specificity, simplicity, and reproducibility for IL-6 determination.
- The technique is suitable for IL-6 quantification in serum samples.
- The integration of fluorescence microscopy imaging provides additional visualization capabilities.
Abstract:
Nanoparticle labels conjugated with biomolecules are used in a variety of different assay applications. In this paper, a sensitive fluoroimmunoassay for recombinant human interleukin-6 (IL-6) with the functionalized Rubpy-encapsulated fluorescent core-shell silica nanoparticles labeling technique has been proposed. IL-6 was measured based on the specific interaction between captured IL-6 antigen and functionalized fluorescent core-shell nanoparticles-labeled anti-IL-6 monoclonal antibody. The calibration graph for IL-6 was linear over the range 20-1250 pg ml(-1) with a detection limit of 7 pg ml(-1) (3 sigma). The regression equation of the working curve is I(F)=7.665+32.499[IL-6] (ng ml(-1)) (r=0.9980). The relative standard deviation (R.S.D.) for 11 parallel measurements of 78 pg ml(-1) IL-6 was 3.2%. Furthermore, the application of fluorescence microscopy imaging in the study of the antibody labeling and sandwich fluoroimmunoassay with the functionalized fluorescent core-shell silica nanoparticles was also explored. This proposed method has the advantage of showing the specificity of immunoassay and sensitivity of fluorescent nanoparticle labels technology. The results demonstrate that the method offers potential advantages of sensitivity, simplicity and reproducibility for the determination of IL-6, and is applicable to the determination of IL-6 in serum samples and enabling fluorescence microscopy imaging for the determination of IL-6.
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