کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5522117 | 1545672 | 2016 | 11 صفحه PDF | دانلود رایگان |
- A comprehensive comparison of 9 immunoassay technologies and 4 cytokine assays
- Assay sensitivity for endogenous not recombinant protein that matters most
- Frequency of endogenous analyte detection (FEAD) is a useful measure of sensitivity.
- Correlation analysis in the absence of gold standard is critical to validate FEAD.
- Performance of IL-2, IL-17a, IL-6, and TNFα across multiple platforms summarized.
A comprehensive cross-platform and cross-assay evaluation using nine technology platforms and four cytokine immunoassays (IL-6, TNFα, IL-17a, IL-2) was performed by comparing assay precision, sensitivity, parallelism and data correlation between platforms. The precision was acceptable for most evaluated assays. In addition to comparing the analytical assay sensitivity using a spiked recombinant analyte in buffer, forty serum samples from both normal controls and multiple sclerosis patients were used to measure the frequency of endogenous analyte detection (FEAD) as a parameter of each assay's ability to detect the endogenous analyte. The highest FEAD measurements were observed on the Simoaâ¢, Erenna®, Milliplex® and Imperacer® platforms. However, only Simoa and Erenna results showed a high correlation across all evaluated cytokine assays, followed by a more moderate correlation of results across platforms for the V-plexâ¢, high sensitivity ELISA and the Ella⢠IL-6 and TNFα assays. In contrast, results from the evaluated cytokine assays on the Milliplex, AMMP⢠ViBE® and Imperacer platforms did not correlate to each other nor to other evaluated assays. Acceptable parallelism was observed for the Simoa, Erenna, V-plex and Ella assays but not for the Milliplex, AMMP ViBE and Imperacer assays. In conclusion, the Simoa, Erenna,V-plex and Ella platforms performed well in one or more evaluated cytokine assays. Among those, the Simoa and Erenna assays had the highest sensitivity for detection of cytokines present at sub-pg/mL levels in human serum. In addition, the cross-platform and cross-assay comparisons demonstrated that different immunoassays may yield different results, which underscores the importance of performing such comparative evaluations, especially in the absence of reliable reference standards for the quantitative assessments of biomarkers in immunoassays.
Journal: Journal of Immunological Methods - Volume 437, October 2016, Pages 53-63