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Updated: Feb 23, 2026
Development and Validation of an Ultrasensitive Single Molecule Array Digital Enzyme-linked Immunosorbent Assay for Human Interferon-α
Published on: June 14, 2018
SERS-based Immunoassay in a Microfluidic System for the Multiplexed Recognition of Interleukins from Blood Plasma:
Agnieszka Kamińska1, Katarzyna Winkler2, Aneta Kowalska2
1Institute of Physical Chemistry, Polish Academy of Sciences, Kasprzaka 44/52, 01-224, Warsaw, Poland. akamin@ichf.edu.pl.
Insights
This study introduces a novel microfluidic device using SERS-active nanostructures for rapid, multiplexed detection of Type 1 cytokines (interleukins IL-6, IL-8, IL-18) in blood plasma. The method offers improved detection limits compared to ELISA, demonstrating high sensitivity and reproducibility.
Area of Science:
- Biomedical Engineering
- Analytical Chemistry
- Nanotechnology
Background:
- Accurate and rapid monitoring of Type 1 cytokines is crucial for diagnosing and managing inflammatory diseases.
- Existing methods like ELISA can be time-consuming and lack the multiplexing capability for simultaneous analysis of multiple cytokines.
- Development of sensitive and specific biosensing platforms is needed for early disease detection.
Purpose of the Study:
- To develop a novel Surface-Enhanced Raman Spectroscopy (SERS)-based microfluidic device for rapid and multiplexed quantification of Type 1 cytokines (IL-6, IL-8, IL-18) in human blood plasma.
- To demonstrate the feasibility of using distinct Raman reporter molecules and multivariate statistical analysis for simultaneous detection of multiple analytes.
- To compare the performance of the developed SERS method with conventional ELISA in terms of sensitivity and detection limits.
Main Methods:
- Fabrication of SERS-active nanostructures integrated into a microfluidic chip.
- Functionalization of nanoparticles with specific antibodies and distinct Raman reporter molecules (DTNB, FC, p-MBA).
- Application of multivariate statistical analysis (Principal Component Analysis - PCA) for signal discrimination and data interpretation.
- Multiplexed detection of IL-6, IL-8, and IL-18 in blood plasma samples.
Main Results:
- Successful development of a microfluidic device enabling rapid and multiplexed detection of IL-6, IL-8, and IL-18.
- Demonstration of PCA's effectiveness in segregating antigen-antibody complexes based on Raman reporter signals.
- Achieved low detection limits: 2.3-4.2 pg·ml⁻¹ (parallel) and 3.8-5.2 pg·ml⁻¹ (simultaneous multiplexed) for the target interleukins.
- The SERS method exhibited superior sensitivity and lower detection limits compared to standard ELISA.
Conclusions:
- The developed SERS-based microfluidic platform provides a sensitive, reproducible, and rapid method for multiplexed cytokine analysis in biological samples.
- This approach represents a significant advancement for point-of-care diagnostics and personalized medicine, particularly for Type 1 inflammatory conditions.
- The study highlights the potential of SERS and multivariate analysis for simultaneous quantification of multiple biomarkers in complex matrices like blood plasma.
Abstract:
SERS-active nanostructures incorporated into a microfluidic device have been developed for rapid and multiplex monitoring of selected Type 1 cytokine (interleukins: IL-6, IL-8, IL-18) levels in blood plasma. Multiple analyses have been performed by using nanoparticles, each coated with different Raman reporter molecules: 5,5'-dithio-bis(2-nitro-benzoic acid) (DTNB), fuchsin (FC), and p-mercatpobenzoic acid (p-MBA) and with specific antibodies. The multivariate statistical method, principal component analysis (PCA), was applied for segregation of three different antigen-antibody complexes encoded by three Raman reporters (FC, p-MBA, and DTNB) during simultaneous multiplexed detection approach. To the best of our knowledge, we have also presented, for the first time, a possibility for multiplexed quantification of three interleukins: IL-6, IL-8, and IL-18 in blood plasma samples using SERS technique. Our method improves the detection limit in comparison to standard ELISA methods. The low detection limits were estimated to be 2.3 pg·ml-1, 6.5 pg·ml-1, and 4.2 pg·ml-1 in a parallel approach, and 3.8 pg·ml-1, 7.5 pg·ml-1, and 5.2 pg·ml-1 in a simultaneous multiplexed method for IL-6, IL-8, and IL-18, respectively. This demonstrated the sensitivity and reproducibility desirable for analytical examinations.
