Luminex^® Metabolic High Performance Panel

Metabolic biomarker sensitivity and precision is shown in Table 1. Assay precision includes the intra-assay coefficient of variation (CV) and inter-assay CV. The intra-assay CV and inter-assay CV are an estimate of variability within and between assays respectively. Assay CV should be less than 15%. As seen in table 1, the R&D Systems metabolic panel meets this precision standard, indicating that the panel is consistent and reproducible.

Table 1. List of metabolic biomarkers, sensitivity, and precision.

To test the new R&D Systems Luminex Metabolic panel, we quantified serum and plasma metabolic biomarkers in 10 people that fasted for 12 hours. One hour after a meal, we quantified post-prandial plasma and serum analyte changes in the same people. As expected, post-prandial (1 hour) levels of C-Peptide and Insulin were elevated in serum and plasma from females (Figure 2) and males (Figure 3). Conversely, Ghrelin levels declined in post-prandial serum and plasma from females and males (Figure 4).

Avoiding false-positive data is key for assay accuracy and specificity. We eliminate false-positive data by adhering to a rigorous quality control process. We also optimize diluents so that analytes on the panel are compatible with each other.

Circulating IL-6 levels should be around 1.8 pg/ml⁴. The R&D Systems Metabolic panel IL-6 measurement is close to the expected value, while the competitor assay is artifactually inflated (Table 2). 

Table 2. Circulating IL-6 data from R&D Systems and competitor Luminex assays. ND=not detectable

Serum Separating tube (SST) serum and EDTA plasma samples were collected from 10 donors for all experiments.  The initial blood draw was collected after 12 hours of fasting.  The donors were then fed a mixed meal and told to return 1 hour after finishing the meal for a second blood draw.  SST samples were injected with Dipeptidyl Peptidase-4 (DPP4) inhibitors immediately after collection then allowed to sit at room temp to clot prior to centrifugation.  Normal EDTA samples were injected with DPP4 inhibitors immediately after collection then placed on ice until centrifugation.  Both the SST serum and normal EDTA samples were separated from the blood cells and Protease Inhibitor Cocktail I (Tocris Catalog number 5500) was added to a 1X concentration.  P800 EDTA samples (BD Biosciences catalog number 366420) were drawn and placed on ice.  No additional inhibitors were added to these tubes, as they contain a proprietary protease inhibitor cocktail. 

Here we have demonstrated that the R&D Systems Luminex Human Metabolic Panel can be used to quantify metabolic biomarkers in human plasma and serum. This assay allows for the simultaneous quantification of 12 metabolic biomarkers in each 50 ul sample.  Our Luminex assay is both faster and provides superior performance to competitor Luminex assays.

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