Abstract
The need for a rapid screening assay for protein expression patterns has led us to develop an antibody array. Samples from rat models of vasculitis (glucan induced granulomatous and immune complex induced dermal Arthus) were tested. An antibody array with 34 antibodies to rat proteins was utilized to compare serum protein levels of injured vs. control animals from these two models. Lower limits of detection were in the upper pg/ml range for many of the tested analytes. Capture antibodies were spotted onto slides and serum proteins detected with biotin-conjugated secondary antibodies and signal generated with streptavidin conjugated Alexafluor. A standard curve for each analyte was generated with recombinant protein standards and used to calculate analyte concentrations. In the glucan injury model there was a statistically significant increase in glucan injury vs. the control groups for ICAM-1 (p<0.01) and CINC-1, IL-12 and Notch-2 (p<0.05, respectively). In the dermal Arthus model we found significantly higher concentrations of CINC-2 & IFN-γ (p<0.05) and IL-1 α & PDGF-AA (p<0.05) in the injured vs. control rats. Findings were verified with classic ELISA technology. These findings demonstrate that multi-antigen based antibody arrays perform well for analysis of serum samples from vasculitis injury models, allowing rapid screening with small sample size. Supported by Pfizer Global Research and Development.
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