A reproducible and sustainable two dimensional HPLC method that removes highly and moderately abundant plasma proteins stepwise and continuously

摘要

Affinity based removal of abundant plasma proteins has been shown to effectively facilitate the quantification of low abundant plasma proteins. In particular, stepwise affinity based removal scheme where highly abundant and moderately abundant proteins are depleted sequentially with separate antibody columns has uncovered even more proteins for biomarker discovery. Though effective in extending the dynamic range of proteomic analysis, the stepwise depletion normally requires two separate LC runs with a concentration step in between which lowers the throughput and increases the variability of the analysis. Here we report an alternative, streamlined HPLC method that removes highly and moderately abundant proteins continuously with two in line affinity-depletion columns. The sustainability and reproducibility of the system were greatly enhanced as a result.