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Updated: Jun 20, 2026
Development and Validation of an Ultrasensitive Single Molecule Array Digital Enzyme-linked Immunosorbent Assay for Human Interferon-α
Published on: June 14, 2018
Neutralizing antibodies against IFN beta in patients with multiple sclerosis: a comparative study of two cytopathic
1Research Laboratory, Hospital Civil, Pab. 5, Sótano, 29009 Málaga, Spain. begoliver@gmail.com
Insights
Neutralizing antibodies (NABs) against interferon-beta (IFN-β) can be reliably measured using either the HEP2/VSV or A549/EMCV bioassays. These methods demonstrate comparable sensitivity and correlate well, ensuring consistent results across laboratories.
Area of Science:
- Immunology
- Virology
- Cell Biology
Background:
- Measuring neutralizing antibodies (NABs) against interferon-beta (IFN-β) is crucial for specialized diagnostics.
- The cytopathic effect (CPE) inhibition bioassay is the gold standard, but variations hinder inter-laboratory comparisons.
- Standardized protocols, like the WHO's A549/EMCV assay, aim to improve result consistency.
Purpose of the Study:
- To compare the sensitivity and correlation of two IFN-β NAB bioassays: HEP2/VSV and A549/EMCV.
- To validate previous laboratory findings obtained using the HEP2/VSV assay.
- To assess if using different cell lines and viruses impacts assay sensitivity.
Main Methods:
- Comparative analysis of two CPE inhibition bioassays: HEP2/VSV and A549/EMCV.
- Calibration of the A549/EMCV assay using a reference IFN-β standard.
- Statistical assessment of association and correlation between the results from both bioassays.
Main Results:
- Both the HEP2/VSV and A549/EMCV bioassays demonstrated comparable sensitivity in measuring IFN-β NABs.
- A significant association and correlation were found between the results obtained from the two different bioassay systems.
- The results confirm that past findings using the HEP2/VSV assay are comparable to those from the A549/EMCV assay.
Conclusions:
- The HEP2/VSV bioassay is as sensitive as the WHO-recommended A549/EMCV assay for measuring IFN-β NABs.
- Established results using the HEP2/VSV assay can be reliably compared with results from the A549/EMCV assay.
- This comparability facilitates consistent IFN-β NAB measurements across different laboratories and research settings.
Abstract:
Neutralizing antibodies (NABs) against IFN beta should be measured in specialized laboratories, using a test of inhibition of the cytopathic effect (bioassay or CPE test), based on the capacity of IFNss to block the infection of live monolayer-cultured cells by a virus, depending on the presence or absence of NABs. The European Federation of Neurological Societies (EFNS) considers this assay to be the gold standard. However, the various different ways to perform this assay complicate comparison of the results between laboratories. The World Health Organization (WHO) has published several recommendations to perform this assay using the A549 cell line and the murine encephalomyocarditis virus (EMCV). In order to validate the results previously obtained in our laboratory with HEP2/VSV, we undertook a comparative analysis of the two bioassays, HEP2/VSV and A549/EMCV, to assess whether the use of different cell lines and viruses influences sensitivity. We also calibrated the A549/EMCV assay with a reference IFNss. Our results confirm that the bioassay with HEP2/VSV is as sensitive as the assay with A549/EMCV and that a significant association and correlation exist in the results between both assays. Thus, past results with HEP2/VSV in our laboratory could be comparable with those obtained with A549/EMCV in both our laboratory and others.
