Evaluation of highly sensitive immunoassay technologies for quantitative measurements of sub-pg/mL levels of cytokines in human serum
| Autor: | Douglas Donaldson, Alvydas Mikulskis, David Yeung, Chase Shen, Shobha Purushothama, Shawn Ciotti, Geoffrey Kuesters, Brian Schlain, Elham Gharakhani |
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| Rok vydání: | 2016 |
| Předmět: |
Serum
0301 basic medicine Analyte Immunology Data correlation Enzyme-Linked Immunosorbent Assay Sensitivity and Specificity 01 natural sciences 03 medical and health sciences medicine Humans Immunology and Allergy Reference standards Observer Variation Chromatography medicine.diagnostic_test Chemistry Ligand binding assay 010401 analytical chemistry Assay sensitivity Reference Standards Serum samples 0104 chemical sciences Highly sensitive 030104 developmental biology Immunoassay Cytokines |
| Zdroj: | Journal of Immunological Methods. 437:53-63 |
| ISSN: | 0022-1759 |
| Popis: | 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 Simoa™, 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-plex™, 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. |
| Databáze: | OpenAIRE |
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