S&S® Serum Biomarker Chip Displays Specificity & Reproducibility for 120 Different Human Biomarker Profiles
Abstract
We introduce a unique tool for the determination of relative abundances for human serum biomarkers. Conceptually similar to DNA microarrays, the S&S® Serum Biomarker Chip (SBC) is a single capture antibody array that was developed for comparative analysis of serum samples in order to identify differences or similarities in protein expression profiles.
The SBC array is manufactured on a nitrocellulose two-pad FAST® slide in which each pad contains 120 antibodies specific for circulating tumorrelated proteins printed in triplicate and arranged as four quadrants per pad. The Universal Linkage System (ULS™) protein labeling kit was employed to label serum proteins with either biotin or fluorescein prior to incubation with the SBC array.
Data from the array was collected using an Axon scanner to detect fluorescent emitted from Dyomics dyes DY™647 and DY™547 conjugated to Streptavidin and anti-Fluorescein antibody respectively. We show that fluorescently labeled biomarker proteins bind specifically to their capture antibody as determined via competition experiments.
The system was tested for reproducibility by running eleven individual experiments and measuring the variation in the signal intensity of each target biomarker. The results show that the CV is less than 20% for 105 of the 120 capture biomarker antibodies.
