IL-2 GoStix Plus

Timelines associated with commonly used methods for measuring cytokines.

Timelines associated with commonly used methods for measuring cytokines. GoStix Plus: a lateral flow-based quantitation method. Fast ELISA: measurement using targeted, preformulated ELISA reagents. Standard ELISA: measurement using standard direct or indirect sandwich assay. Bead array: multiparametric measurement by flow cytometry of bead-based, immunocapture of cytokines. Glass array: multianalyte measurement using quantitative, glass-slide multiplex ELISA microarray platform. Assay durations were taken from manufacturers’ protocols.

GoStix Plus assays provide results similar to ELISA.

GoStix Plus assays provide results similar to ELISA. Purified, human primary CD3+ T cells were activated for 24 hours using tetrameric antibody complexes to cell-surface ligands CD3, CD28, and CD2. Cell culture supernatant was collected at the time points indicated post-activation and analyzed using both IL-2 GoStix Plus and an ELISA. For the ELISA samples, several dilutions of each sample were run on the assay as suggested by the manufacturer.

Comparison of T-cell activation methods using IFN-γ GoStix Plus and ELISA-based methods.

Comparison of T-cell activation methods using IFN-γ GoStix Plus and ELISA-based methods. Human primary T cells were isolated from peripheral blood (PB) mononuclear cells (MNCs) using negative selection. T cells were subsequently activated with either CD3/CD28-coated beads, CD3/CD28 tetramer complexes, or PHA (10 μg/ml). Supernatant samples were collected at the indicated timepoints and assayed for IFN-γ protein using IFN-γ GoStix Plus or an IFN-γ ELISA. Assay results were converted into IU/ml values using NIBSC reference standard 82/587.

Cytokine GoStix Plus workflow.

Cytokine GoStix Plus workflow. Apply 20 µl of sample to a GoStix lateral flow cassette followed by 80 µl of chase buffer. Wait 10 minutes for the test to run, then scan the cassette using the GoStix Plus smartphone app.

Quantitation of human TH1 cytokines using GoStix Plus lateral flow assays.

Quantitation of human TH1 cytokines using GoStix Plus lateral flow assays. Purified, human primary CD3+ T cells were activated for 24 hours using tetrameric antibody complexes to cell-surface ligands CD3, CD28, and CD2. Cell culture supernatants were collected at the indicated timepoints post activation and analyzed using IL-2, IFN-γ, and TNF-α GoStix Plus assays and the accompanying GoStix Plus app.