Transition from identity to bioactivity‐guided proteomics for biomarker discovery with focus on the PF2D platform

Abstract

Proteomic strategies provide a valuable tool kit to identify proteins involved in diseases. With recent progress in MS technology, high throughput proteomics has accelerated protein identification for potential biomarkers. Numerous biomarker candidates have been identified in several diseases, and many are common among pathologies. An overall strategy that could complement and strengthen the search for biomarkers is combining protein identity with biological outcomes. This review describes an emerging framework of bridging bioactivity to protein identity, exploring the possibility that some biomarkers will have a mechanistic role in the disease process. A review of pulmonary, cardiovascular, and CNS biomarkers will be discussed to demonstrate the utility of combining bioactivity with identification as a means to not only find meaningful biomarkers, but also to uncover functional mediators of disease.

Astrocytic TIMP-1 regulates production of Anastellin, a novel inhibitor of oligodendrocyte differentiation and FTY720 responses

“…The conditioned media was then run through a size filtration column to separate out the proteins by their molecular weight and collected. Eluted proteins were run through protein fractionation 2-dimensional (PF2D), where the proteins were then further separated by a gradient of isoelectric point and then hydrophobicity, as previously described (Beckman Coulter, Pasadena, CA) (34, 58). Identities of proteins differentially produced in TIMP-1 KO and wild-type mixed glial cultures were determined by mass spectrometry (MS/MS).…”