Case Study: Model-to-Man

Enabling optimal and humane study designs through multiplex protein detection in low sample volumes

Mouse

Mice play a pivotal role in disease research and therapy evaluation.

The small biofluid and supernatant volumes that can be non-destructively obtained from animal and cell culture models often impose limitations to study design. Sample pooling obscures individual variability. Repeat or time course measures are done with multiple rather than a single individual, which confounds analysis. Experiment setup becomes complicated and involves high numbers of animals or cultures.

The extreme sensitivity and high multiplexing capacity of LUNARIS™ Technology overcomes these limitations. To demonstrate this robust cytokine quantification at even low sample volumes, blood from four immunized wild-type and four immunized knockout C57BL/6J mice was analyzed on the LUNARIS™ Platform.

LUNARIS reader

The LUNARIS™ Platform is designed for translating insights from model to man.

The LUNARIS™ Mouse 12-Plex Cytokine Kit (LMC-20121) was used to simultaneously assay for IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12 (p70), IL-13, IL-17, TNF-α, IFN-γ and GM-CSF in repeat blood draws 1, 3, 6, and 18 hours after immunization. Readout was performed on the LUNARIS™ Reader (LRS-001) and data were automatically evaluated with the LUNARIS™ Analysis Suite.

All cytokines were measured in 3 µL serum from individual blood draws at each time point. Cytokine levels as low as 20–100 pg/mL were recorded. Data from only three cytokines are shown here.

Data

Fig. 1 Cytokine kinetics in individual immunized wild-type and knockout mice over 18 hours

Absolute IL-10, IL-12 (p70), and IFN-γ serum levels were slightly decreased in knockout (KO) mice compared to wild-type (WT). Both study groups exhibited variability among individuals.

Source: AYOXXA Field Application Specialist Dr. Isabelle Koxholt in collaboration with the Institute of Experimental Immunology Bonn. Poster presented at the Potsdam Proteomic Forum (April 2017.)

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