The PRMM IgG array fabrication with protein G or without protein G and array-based immunoassay procedures.

<p>Six PRMMs were directly labeled with fluorescent dye and probed with antibody slides (aldehyde-derivatized slides coated with or without protein G). Step a: The aldehyde-derivated slide was coated with protein G. Step b: The antibody was printed and immobilized on the protein G-coated slide at 4°C for 2 hours. Step c: The antibody was printed and immobilized on the aldehyde slide at 4°C for 14 hours. Step d: The samples were labeled with fluorescent dye and incubated with the antibody microarray slides at room temperature for 1 hour. Step e: The unbound samples were washed several times. Finally, the signals were directly detected using a fluorescence microarray scanner.</p

Cross-reactivity analysis of the array-based immunoassays with protein G (gray bar) and without protein G (black bar).

<p>Each single PRMM was individually probed with the IgG arrays containing all of the PRMM antibodies: (a) SP, (b) CGRP, (c) NGF, (d) BDNF, (e) TNF-α, and (f) β-endorphin. The error bars represent the standard deviations of four measurements.</p

Sensitivity, dynamic range and LOD for the detection of PRMMs using protein G-facilitated immunoassays.

<p>Note:</p><p>Dynamic range: from LOD to the “saturation point” (beyond which no significant signal increase was observed) of the dose-response curve.</p><p>Sensitivity: (the intensity of saturation point - LOD)/concentration difference.</p