Homebrew simoa assay

The Simoa IFNα assay was developed using a Quanterix Homebrew Simoa assay according to the manufacturer’s instructions and using two autoantibodies specific for IFNα isolated and cloned from two APS1/APECED (autoimmune polyendocrinopathy candidiasis ectodermal dystrophy) patients (Rodero et al., 2017a (link)). The 12H5 antibody clone was used as a capture antibody after coating on paramagnetic beads (0.3 mg/ml), and the 8H1 was biotinylated (biotin-to-antibody ratio = 30:1) and used as the detector.

The Simoa IFN-α assay (Rodero et al., 2017 (link)) was developed using a Quanterix Homebrew Simoa assay according to the manufacturer’s instructions, and using two autoantibodies specific for IFN-α isolated and cloned from two APS1/autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy patients as described (Meyer et al., 2016 (link); Rodero et al., 2017 (link)). The 8H1 antibody clone was used as a capture antibody after coating on paramagnetic beads (0.3 mg/ml), and the 12H5 was biotinylated (biotin/antibody ratio = 30/1) and used as the detector. Recombinant IFNα17/αI (PBL Assay Science) was used to produce a standard curve after cross-reactivity testing. The limit of detection was calculated as the mean value of all blank runs +3 SD and was 0.23 fg/ml.