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Selection of Standards for Bio-Plex Cytokine Assays, Rev A

ng the following formula: (observed concentration) / (expected concentration) x 100. If the regression analysis is accurate, the calculated concentrations will be very close to the actual concentrations of the standards. This method yields information about the dynamic range, sensitivity, and accuracy of calculated concentrations of an assay (Baud 1993). The recovery value for each standard is an indication of the accuracy of the calculated concentration. A value within the range of 70130% is considered accurate recovery. The dynamic range of the assay, or the range where accurate concentrations may be quantitated, is therefore indicated by the range of standards yielding a recovery of 70130%.

A comparison of the standards recovery using broad range and high-sensitivity standards for three different cytokines is shown in Tables 2 and 3. The broad range standard concentrations where recovery is 70130% is 1.95 to 8,000 or 32,000 pg/ml, depending on the cytokine assayed. The high-sensitivity standards yield a narrower dynamic range of 0.2 or 0.78 to 3,200 pg/ml. In all three assays, the broad range standards enable the quantitation of a much wider range of concentrations.

A general indication of assay sensitivity is given by the lowest concentration in a set of standards that yields accurate recovery (70130%). Since the lowest concentration of the high-sensitivity standards is lower than the broad range standards (i.e., 0.2 pg/ml vs. 1.95 pg/ml, respectively), it would be expected that the high-sensitivity standards would yield better recovery at a lower concentration, and that the high-sensitivity standards should be chosen to maximize sensitivity in an assay. Tables 2 and 3 illustr
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