The SBA was produced by covalently coupling Abs to MagPlex Beads (Luminex), which are magnetic, bar-coded beads where each bar code was defined by a unique combination of infrared and near-red dyes. Each Ab was coupled to a unique bead identity as previously described (17). In brief, 1.6 μg of each Ab was diluted in MES buffer [100 mM 2-(N-morpholino)ethanesulfonic acid (pH 5.0)] to a final volume of 100 μl. The carboxylated surface of the magnetic beads was then activated with N-hydroxysuccinimide (Pierce) and 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (ProteoChem). After 20 min of incubation with the activation solution, the diluted Abs were added to the beads, and coupling occurred for 2 hours at room temperature (RT). Beads were then washed and stored in a protein-containing blocking buffer, BRE (Roche), with the addition of ProClin 300 (Sigma-Aldrich). Coupling efficiency for each Ab was determined by incubating the beads with PE-conjugated anti-rabbit IgG or PE-conjugated anti-mouse IgG (Jackson ImmunoResearch).

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