Multiplexed Analysis of Circulating Prostate Tumor and Host Response Markers

摘要

During the second year of this project we have made further strides toward the broad goals of this project to develop improved blood-based methods for prostate cancer diagnostics and to better understand the nature of the changes in the blood of prostate cancer patients. The first year of the project was used to lay some of the fundamental foundations for the later work: we answered valuable questions about the optimal strategies for using antibody microarrays, we better characterized the performance of the antibodies, and we significantly improved the experimental protocols. The work of the second year has built upon those developments and has resulted in several accomplishments. We further developed and improved the methods by instituting improved quality control for the antibodies, by developing an improved ability to run samples in high-throughput, and by identifying a better-performing microarray surface. We performed initial profiling studies on serum samples from prostate cancer patients and controls using antibody microarrays containing 96 or 91 antibodies. The preliminary analyses identified areas of the technology that needed improving or that required an alternate strategy. We have addressed and solved the potentially confounding influence of species-dependent, non-specific binding, and we are Using the alternate strategy of sandwich immunoassays for a select set of low-abundance analytes. The early analyses also identified several interesting differences between the patient groups, which will be farther investigated. Our improved methods are currently being used for comprehensive profiling studies of our serum sample sets, and the data from these studies will be analyzed to fulfill the goals of year 3.