Fc Array Analysis of Polyclonal Antibody Responses

Fc array analysis was performed blinded to group as described (47 (link), 48 (link)) to evaluate polyclonal Ab responses. Briefly, plasma was centrifuged for two min at 14.8 x g, then aliquoted and analyzed for binding to a panel of antigens listed in Supplementary Table 1. Plasma was diluted 1:1,000 for detection reagents (Supplementary Table 2) of tetramerized Fcγ receptors (Source: Duke Protein Production Facility) and anti-rhesus IgG (Source: Southern Biotech). For increased sensitivity, detection reagents C1q (Source: Sigma) and anti-human IgA (Source: Southern Biotech) were collected with a plasma dilution of 1:250, along with a replicate of the anti-rhesus IgG. Data collection was performed using Luminex Exponent version 4.2 software.