Figure 2. Stage II Design, Analysis and Customized Arrays.

In Stage II focused arrays were analyzed (a), including multiple normalization steps, and analysis of variance (ANOVA) to ascertain statistical significance in obtaining a high-priority candidate set of three proteins. Each custom array contained 12 blocks, such as the example (b), and was scanned for Immunoglobulin G, IgG, reactivity [using wavelength 532 nm (green) Laser Emission Fluorescence] and protein levels [GST fusion tag expression levels using wavelength 635 nm (red) Laser Emission Fluorescence], with the superposition of both green and red emission spectra from scanning as shown. The protein arrays from patient signals (c) show increased reactivity to IgG as compared to a negative control. Black and low luminosity indicate no and lower reactivity respectively.