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. 2016 Oct 17;1(4):586–599. doi: 10.1021/acsomega.6b00188

Figure 6.

Figure 6

Six-plex ELISA titrations as a function of target size and percentage agarose in the beads show the effect of agarose concentration on bead pore size (pore size 1 = 4 wt % agarose, pore size 2 = 2 wt % agarose and pore size 3 = 1 wt % agarose; lower agarose percentage gives larger average pore size). An IgG Ab against GM-CSF Ag is attached to each of the three different pore size beads and an IgG Ab against IgM Ag is likewise attached to each of the three different pore size beads. The six pore size–capture probe combinations, each of which has its own unique bead optical code, are pooled together and titrated with increasing concentrations of either 15–30 kDa GM-CSF antigen (top) or ∼970 kDa IgM (bottom). The beads are treated with a biotinylated monoclonal Ab against the target species and then stained with SAPE. The top plot shows that none of the GM-CSF titrant is captured by the three codes with the anti-IgM capture probes, but the GM-CSF is taken up by the anti-GM-CSF capture Ab bead codes with larger pores capturing more protein target. The bottom plot shows that none of the IgM titrant reacts with either the bead codes with the anti-GM-CSF capture Ab or the small-pore 4 wt % agarose beads with anti-IgM capture Abs, but the large IgM target is taken up by the larger pore (1 and 2% agarose) beads.