B) Schematic illustration of the SPR-sensor chip flowcell. A schematic illustration of the flowcell and its detection system is shown where anti-IgA antibodies are circulated through the flowcell and as binding to, and dissociation from, the immobilised samples occur the change of refraction is monitored by the detector and recorded.
C: Recorded binding curves from the first set of three analytes. A total of 400 simultaneously recorded binding curves from label free analysis with anti-IgG, anti-HSA, and running buffer. Both anti-IgG and the positive control anti-HSA show strong binding to all spots except the sample buffer spot. Results from anti-IgG show no correlation to results from anti-IgA indicating that the IgA-deficient samples contains normal IgG levels. Anti-HSA functions as a positive control and indicates that all spots are present.
D) Recorded binding corves from the second set of three analytes. Binding curves from label free analysis with negative control (anti-rabbit), anti-IgA, and anti-C3 as a second positive control. They show minimal binding of the negative control to all spots and varying binding from anti-IgA, which is to be expected. The Spearman correlation coefficient between replicate chips was R = 0.98 which implies that single chip would be enough in order to validate the results from the fluorescent arrays.
