Evo 150

The 9827 selected SNPs were submitted to the Illumina Assay Design Tool for design score calculation (www.illumina.com) and 8580 of them were successfully synthesized by Illumina manufacturing processes. The SNP genotyping assay was performed on an Illumina^® Infinium HD iSelect Custom Genotyping Array according to the standard Illumina's protocol, using 200 ng of genomic DNA per sample. The extension and staining steps were operated on a Tecan Evo‐150 liquid‐handling robot. Fluorescence intensities were read with Illumina^® iScan Control software and allele calling was carried out using the Genome Studio v2.0 software (Illumina Inc., San Diego, CA, USA). All data were visually inspected and manually re‐scored if any errors were evident in the calling of the SNP clusters by the default algorithm. Reproducibility error rates were calculated between the control sample and SNP replicates.

Crude EDTA plasma was stored at -80°C. Prior to aliquoting, samples were transferred to -20°C overnight, thawed at 4°C and then vortexed and centrifuged at 3000 rpm for 2 minutes (Allegra X-12, Beckman Coulter). Using a liquid handling robot (EVO150, TECAN), samples were randomized across 96-well microtiter plates (Supplementary Information). Protein labelling was performed with biotin, as previously described by Drobin et al. [11] (link) Briefly, EDTA plasma was diluted ~1:10 in Phosphate buffered saline (PBS) (09-9400-100, Medicago) and biotinylated with EZ-Link-NHS-PEG4-Biotin (21330, Thermo Scientific) dissolved in dimethyl sulfoxide (DMSO) (276855, Sigma-Aldrich). Following 2 h incubation at 4°C, the reaction was quenched with Tris-HCl 0.5 M, pH 8.0. Prior to analysis, a test was performed to confirm successful biotinylation: 24 randomly selected samples were diluted, incubated with the antibody array, and analyzed using the protocol detailed below (see Antibody suspension bead array assays). Further, a test measuring reactivity of human IgM to rabbit IgGs was performed as described in the Supplementary Information and Materials and Methods (Fig. S11). Labelled plasma was stored at -20°C until analysis.

For a high-throughput 384-well protocol, we used the Biotek EL406 automated plate washer and the Tecan EVO150 liquid hander robot, as described in our previous paper (39 (link)). This paper also describes the PEIMAX high-throughput transfection details for HEK293T cells.