In:
The Journal of Immunology, The American Association of Immunologists, Vol. 198, No. 1_Supplement ( 2017-05-01), p. 61.6-61.6
Abstract:
Most immunoassays quantify cytokines with pg/mL sensitivity, which is insufficient to detect those present at low levels. MSD’s next-generation S-PLEXSM assay format was developed to quantify cytokines with fg/mL sensitivity. Here we describe measuring IL-2, IL-4, IL-6, IL-10, and IL-17A levels in normal sera and the supernatants of model cell lines using S-PLEX assays. The assays demonstrated a dynamic range of approximately four orders of magnitude. Standard intra-plate coefficients of variation (CVs) ranged from 3%-15% and inter-plate CVs ranged from 8%–18%. The lower limit of detection (LLOD) was & lt;1 fg/mL for all assays except IL-17A (11 fg/mL). IL-2, IL-6, IL-10, and IL-17A were detectable in 100% of normal sera samples (n=36–75) and IL-4 was detectable in 40% of normal sera samples (n=75). The average concentrations were & lt;1 pg/mL for normal samples. To confirm the sensitivity of these assays and their ability to detect native analytes, we characterized a panel of 23 cell lines that are models for cytokine secretion. As an example, the MOLT-4 cell line derived from acute lymphoblastic leukemia natively expressed detectable levels of IL-2 (1,031 fg/mL), IL-4 (221 fg/mL), IL-10 (60 fg/mL), and IL-17A (30 fg/mL). The expression profiles of these cell lines confirmed the sensitivity of the S-PLEX assays. S-PLEX cytokine assays provide a 100 to 1,000-fold lower LOD than standard ELISAs, allowing the determination of baseline cytokine levels in many sample types.
Type of Medium:
Online Resource
ISSN:
0022-1767
,
1550-6606
DOI:
10.4049/jimmunol.198.Supp.61.6
Language:
English
Publisher:
The American Association of Immunologists
Publication Date:
2017
detail.hit.zdb_id:
1475085-5
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