pr049776f_si_001.xls (1.53 MB)
Download fileDetermination of Relative Protein Abundance by Internally Normalized Ratio Algorithm with Antibody Arrays
dataset
posted on 2005-06-13, 00:00 authored by Oskar Andersson, Mark Kozlowski, Tatiana Garachtchenko, Corina Nikoloff, Nancy Lew, David J. Litman, Grigoriy ChagaIn this paper, we report an experimental setup and mathematical algorithm for determination of relative
protein abundance from directly labeled native protein samples applied to an array of antibodies. The
application of the proposed experimental system compensates internally at each array element for a
number of deficiencies in array experiments such as differential labeling efficiency in dual color assay
systems, differential solubility of protein molecules in dual color assay systems, and differential affinity
of capture reagents toward proteins labeled with two different fluorescent dyes. This system offers full
compensation for variable amounts of capture reagents on separate array structures, as well as limited
compensation for nonspecific interactions between capture reagents and analytes. The proposed
experimental strategy enables the use of a large number of capture reagents to develop a true multiplex
analysis system that will yield complete relative protein abundance information in two biological
systems.
Keywords: antibody array • dual color detection • protein abundance • multiplex protein analyses • internally
normalized ratio