Multiplex Enzyme-Linked Immunosorbent Assay (ELISA) for Detection of Human Cytokines in Culture Supernatants

Multiplex Enzyme-Linked Immunosorbent Assay (ELISA) for Detection of Human Cytokines in Culture Supernatants

Standard Number ITA-27
Organization Nanotechnology Characterization Laboratory USA
Level National
Category Test Method | Characterization | Measurement
Status
  • SEP 2020 Published
ABSTRACT
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. 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 (IFNα, IFNβ, IFNω), type ΙΙ IFN (IFNγ), type III IFN (IFNλ), 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 allowssimultaneous analysis of a broad spectrum of markers.