Quantitation of reduced IL-33 levels in human serum: mitigating interference from endogenous binding partners
IL-33 is a potential therapeutic target but commercially available assays for the quantitation of systemic IL-33 have poor reliability. In commercial IL-33 kits, interference from endogenous binding partners (e.g., soluble ST2) causes under-quantitation. Mitigating this required acid dissociation an...
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Veröffentlicht in: | Bioanalysis 2021-12, Vol.13 (23), p.1751-1760 |
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Sprache: | eng |
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Zusammenfassung: | IL-33 is a potential therapeutic target but commercially available assays for the quantitation of systemic IL-33 have poor reliability.
In commercial IL-33 kits, interference from endogenous binding partners (e.g., soluble ST2) causes under-quantitation. Mitigating this required acid dissociation and addition of the detection reagent simultaneously with the capture step. This enabled detection of total, reduced (active) levels of IL-33 in human serum (LLOQ 6.25 pg/ml).
Acid treatment of serum samples dissociates IL-33 from endogenous binding partners, increasing soluble ST2 tolerance to >1000 ng/ml. The modified method was specific for reduced endogenous IL-33. Analysis of over 300 samples from individuals with and without asthma and with different smoking status revealed no difference in serum IL-33. |
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ISSN: | 1757-6180 1757-6199 |
DOI: | 10.4155/bio-2021-0172 |