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Mouse IgG gRAD
This example illustrates how gRAD can be used to produce a quantitative rapid assay for mouse IgG concentrations e.g. in culture supernatants.
The test reagent is a solution consisting of biotinylated goat anti-mouse-IgG Fc-specific antibody 20 µg/ml and gold colloid coated with goat anti-mouse-IgG Fab
antibody (OD 0.4 at 540 nm) in running buffer. Standards were produced by diluting a mouse IgG1 stock in running buffer and each standard was tested
in triplicate. 100 µl of test reagent were mixed with 100 µl of the relevant standard and allowed to equilibrate for 5 minutes.
150 µl of the mixture were then applied to the gRAD.
The results are shown below, demonstrating the precision of the test over the range of the standard curve. At analyte
concentrations over 275 ng/ml the curve flattens off to such a degree as to limit the precision of the assay. Values corresponding to analyte concentrations above 275 ng/ml indicate that the sample should be further diluted 1/5 and retested.
Over the range of 8.6 ng/ml to 275 ng/ml the test gave coefficient of variation (CV) values for the calculated concentrations ranging from
6.0% to 8.7%, comparable with ELISA.
Standard curve and table of results obtained as described, showing the values obtained for mouse IgG1 concentrations
ranging from 4.3 ng/ml to 550 ng/ml. The mean, standard deviation (SD) and CV at each analyte concentration are given as well
as the analyte concentrations calculated from the standard curve. The error bars indicate 1 SD from the mean.
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