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MAGPIX limit of detection assays for vaccinia virus.

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posted on 2014-09-11, 03:18 authored by Scott A. Walper, Jinny L. Liu, Daniel Zabetakis, George P. Anderson, Ellen R. Goldman

Top two panels show purified sdAbs used as both captures and tracers in bead-based assays to detect the vaccinia virus. The bottom panel shows the assay using conventional antibodies. Capture antibodies are plotted with corresponding biotinylated (Bt) tracer sdAbs indicated above each graph. The Y-axis is a measure of the signal over background while the X-axis represents the concentration of the vaccinia virus. Insets show a close-up of the lower part of the graph, the blue line indicates the LOD, defined as a signal to background ratio of 3. Limits of detection for best performing sdAb capture-tracer pairs approach 4×105 pfu/ml. Sandwich assays for vaccinia detection were performed on multiple days and reproduced; a representative data set is shown. The MAGPIX assays measures binding from at least 50 beads and the error of the mean fluorescence intensity is within 5%.