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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Bibliographic Details
Published in:Bioanalysis 2021-12, Vol.13 (23), p.1751-1760
Main Authors: Zylstra, Joshua, Partridge, Michael A, Sumner, Giane
Format: Article
Language:English
Subjects:
bioanalysis
biomarker
IL-33
LBA
total assay
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Summary: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.
ISSN:1757-6180
1757-6199
DOI:10.4155/bio-2021-0172