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Updated: Aug 13, 2026
Development and Validation of an Ultrasensitive Single Molecule Array Digital Enzyme-linked Immunosorbent Assay for Human Interferon-α
Published on: June 14, 2018
A time-resolved fluorescence immunoassay (DELFIA) increases the sensitivity of antigen-driven cytokine detection
Gina Allicotti1, Eva Borras, Clemencia Pinilla
1Torrey Pines Institute for Molecular Studies, San Diego, California 92121, USA.
Insights
A new assay, dissociation-enhanced lanthanide fluoroimmunoassay (DELFIA), significantly improves the detection of low cytokine levels from T cells. This enhanced sensitivity allows for more precise quantification of cytokine production in research settings.
Area of Science:
- Immunology
- Biochemistry
- Assay Development
Background:
- Quantifying low levels of cytokines released by T cells is crucial for understanding immune responses.
- Standard enzyme-linked immunosorbent assays (ELISA) may lack the sensitivity required for detecting these low-abundance molecules.
Purpose of the Study:
- To develop and evaluate a novel sandwich dissociation-enhanced lanthanide fluoroimmunoassay (DELFIA) for improved quantification of cytokines.
- To compare the sensitivity of DELFIA with standard colorimetric ELISA for cytokine detection.
Main Methods:
- Development of a sandwich DELFIA assay.
- Comparison of DELFIA with colorimetric ELISA using mouse IL-2 and human GM-CSF assays.
- Assessment of assay sensitivity and sample volume requirements.
Main Results:
- DELFIA demonstrated 8- to 27-fold enhanced sensitivity for mouse IL-2 compared to ELISA.
- DELFIA showed a 10-fold increase in sensitivity for human GM-CSF compared to ELISA.
- The increased sensitivity enables the use of smaller sample volumes and more assays per sample.
Conclusions:
- Sandwich DELFIA is a highly sensitive, nonisotopic method for cytokine quantification.
- DELFIA offers a significant improvement over standard ELISA for detecting low levels of antigen-driven cytokine production.
- This assay is a valuable tool for researchers studying T cell-mediated immune responses.
Abstract:
In an effort to improve the quantification of the low levels of cytokines released in response to antigenic stimulation of T cells, a sandwich dissociation-enhanced lanthanide fluoroimmunoassay (DELFIA) was developed and compared to a standard sandwich ELISA. The DELFIA enhanced the sensitivity of a mouse IL-2 assay 8- to 27-fold, and a human GM-CSF assay 10-fold, as compared to colorimetric ELISA. The increase in sensitivity allows for the use of lower sample volumes per well, and the ability to run more assays per supernatant sample. This sensitive, nonisotopic alternative to other cytokine detection methods will be useful for those researchers wanting to quantitate low levels of antigen-driven cytokine production.
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