ELISA and multiplex technologies for cytokine measurement in inflammation and aging research - PubMed (original) (raw)

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ELISA and multiplex technologies for cytokine measurement in inflammation and aging research

Sean X Leng et al. J Gerontol A Biol Sci Med Sci. 2008 Aug.

Abstract

Over the last decade there has been an enormous expansion of research focused on defining the role of inflammation in aging, age-related diseases, disability, and frailty. The availability of methods to measure cytokines and other inflammatory mediators or markers with high sensitivity and specificity is critically important. Enzyme-linked immunosorbent assay (ELISA), the most widely used and best validated method, is limited by its ability to measure only a single protein in each sample. Recent developments in serum cytokine quantification technology include multiplex arrays, which offer the potential of better evaluating the complexity and dynamic nature of inflammatory responses and offer substantial cost and sample savings over traditional ELISA measurements. Despite potential advantages of this new technology, experience with these techniques is limited, and it has not emerged to date as the gold standard in inflammatory mediator measurement. This article reviews ELISA and the emerging multiplex technologies, compares the cost and effectiveness of recently developed multiplex arrays with traditional ELISA technology, and provides specific recommendations for investigators interested in measuring serum inflammatory mediators in older adults.

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Figures

Figure 1

Figure 1

Schematic illustration of the experimental principle for ELISA and bead-based multiplex assays. Both involve capture antibody and detection antibody specific to the cytokine(s) of interest, offering immunological specificity. Enzyme or fluorochrome linked to the detection antibody is the method of detection with signal amplification, offering sensitivity. Panel A illustrates the basic protocol for traditional double antibody sandwich ELISA. Proprietary bead sets provide additional differential detection power in bead-based multiplex arrays (Panel B).

Figure 1

Figure 1

Schematic illustration of the experimental principle for ELISA and bead-based multiplex assays. Both involve capture antibody and detection antibody specific to the cytokine(s) of interest, offering immunological specificity. Enzyme or fluorochrome linked to the detection antibody is the method of detection with signal amplification, offering sensitivity. Panel A illustrates the basic protocol for traditional double antibody sandwich ELISA. Proprietary bead sets provide additional differential detection power in bead-based multiplex arrays (Panel B).

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