The protein microarray is an emerging application format in diagnostics. High quality microarrays are produced on various substrates.  However, the distribution of printed biomolecules on these substrates is often non-homogeneous, commonly known as the �doughnut or coffee-stain effect�. We studied the influence of pluronic F127 in the printing buffer on spot morphology in porous nitrocellulose membranes (11 �m thick); a high signal-to-noise substrate. Antibodies specific for discriminating tags were printed on nitrocellulose membrane pads on slides using phosphate buffered saline (pH 7.4) and various concentrations of pluronic F127 as a printing buffer. Double-tagged amplicons were sandwiched in an one-step incubation between the printed antibodies and streptavidin conjugated flurophore. Results were analyzed by the 'Z-stack' method of the confocal laser scanning microscope. The intensity distribution in each stack was assessed. Printing antibodies with increasing concentrations of pluronic F127 up to 0.2% (w/v) showed an increasing uniformity of spot morphology.

Influence of pluronic F127 on the distribution of protein molecules and resulting spot morphology of microarrays in a porous nitrocellulose substrate

243rd National Meeting (2012)

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