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Updated: Jul 7, 2026
Development and Validation of an Ultrasensitive Single Molecule Array Digital Enzyme-linked Immunosorbent Assay for Human Interferon-α
Published on: June 14, 2018
Suspension microarrays for the identification of the response patterns in hyperinflammatory diseases
Hsin-Yun Hsu1, Silke Wittemann, E Marion Schneider
1NMI Natural and Medical Sciences Institute at the University of Tübingen, Reutlingen, Germany.
Insights
A new multiplex assay accurately measures 11 soluble receptors in plasma. This method distinguishes sepsis from trauma by identifying specific elevated receptor levels in sepsis patients.
Area of Science:
- Biochemistry
- Immunology
- Clinical Chemistry
Background:
- Multiplex sandwich immunoassays enable simultaneous analysis of multiple analytes.
- Protein arrays are crucial for studying inflammatory diseases by tracking plasma protein changes.
- Bead-based systems like Luminex are established for multiplexed immunoassays.
Purpose of the Study:
- To develop and validate an 11-plex assay for soluble receptor quantification in plasma.
- To analyze plasma soluble receptor concentrations in critically ill patients with trauma or sepsis.
- To identify potential biomarkers differentiating sepsis from trauma.
Main Methods:
- Development of a multiplexed bead-based immunoassay using the Luminex system.
- Quantification of eleven soluble receptors (sTNF-RI, sTNF-RII, sIL-2R, sgp130, sFas, sRAGE, sE-selectin, sICAM-1, sVCAM-1, sMIF-1, sFasL) in 100 plasma samples.
- Statistical analysis to compare receptor levels between trauma and sepsis patient groups.
Main Results:
- The 11-plex assay demonstrated good precision, accuracy, and no cross-reactivity.
- Plasma samples from 36 critically ill patients were successfully analyzed.
- Four molecules (sFas, sICAM-1, sTNF-RI, sTNF-RII) showed significantly higher concentrations in sepsis patients compared to trauma patients.
Conclusions:
- The developed multiplex assay is reliable for analyzing soluble receptor profiles in critically ill patients.
- Elevated levels of sFas, sICAM-1, sTNF-RI, and sTNF-RII can serve as discriminatory biomarkers for sepsis.
- This assay aids in understanding inflammatory disease mechanisms and patient stratification.
Abstract:
Miniaturized and parallelized sandwich immunoassays allow the simultaneous analysis of a variety of parameters in a single experiment. Bead-based protein array systems or suspension microarrays are well-established multiplex sandwich immunoassay formats. To study inflammatory diseases, protein arrays can be used to analyze changes in plasma protein levels, such as cytokines, chemokines, soluble receptors, and matrix metalloproteinases. Using the bead-based Luminex system, multiplexed sandwich immunoassays have been developed to analyze the plasma concentrations of soluble receptors: sTNF-RI, sTNF-RII, sIL-2R, sgp130, sFas, sRAGE, sE-selectin, sICAM-1, sVCAM-1, sMIF-1 and sFasL. This newly established 11-plex soluble receptors assay demonstrated acceptable intra-assay and inter-assay precision, appropriate accuracy, and no crossreactivity between analytes. Using this assay, 100 plasma samples derived from 36 critically ill intensive care unit (ICU) patients with trauma or sepsis were analyzed for their soluble receptor plasma concentrations. Results obtained allowed grouping of patients' samples into a trauma and a sepsis group. Four candidate molecules: sFas, sICAM-1, sTNF-RI, and sTNF-RII had higher concentrations in patients with sepsis than in those with trauma, contributing the highest discriminatory values to define the nature of the inflammatory disease originating from pathogen-involved (sepsis) or pathogen-independent inflammation.
