Cytokines, chemokines and interferons (IFN) are soluble mediators of inflammation and a variety of responses that orchestrate both innate and adaptive immunity in health and disease [1-10]. Understanding the induction of these biomarkers in response to a drug product helps to establish the product’s safety profile and shed light on the mechanism of action or mechanism of toxicity. This document describes experimental procedure for analysis of culture supernatants by multiplex ELISA to detect presence of type I IFNs (IFNa, IFNb, IFNw), type II IFN (IFNg), type III IFN (IFNl), cytokines and chemokines (IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p70, IL-13, IL-15, IL-17, IL-21, IL-22, IL-23, IL-27, IP-10, MCP-1, MCP-2, MIP-1α, MIP-1β, RANTES, TNFα) in culture supernatants. NCL protocol ITA-10 should be referred to for details of preparation of culture supernatants. Protocols for single-plex ELISA (ITA-22, ITA-23, ITA-24 and ITA-25) can also be considered. The advantage of this protocol over single-plex ELISA is that it allows simultaneous analysis of a broad spectrum of markers.
Researchers should cite this work as follows: