Using SELDI-TOF Mass Spectrometry on Amniotic Fluid and for Clinical Proteomics and Theranostics in Disorders of Pregnancy

摘要

Clinical proteomics encompasses a multitude of experimental approaches, tools and techniques based on proteomics technology that are directly aimed to accelerate and improve diagnosis and treatment of human diseases. Surface-enhanced laser desorption ionization time-of-flight (SELDI-TOF) mass spectrometry is a variant of matrix-enhanced laser desorption ionization (MALDI) that makes use of chemically-modified surfaces to reduce the complexity of biological samples prior to separation in the mass analyzer. Compared to other proteomic techniques, SELDI has several important advantages such as ability to analyze complex biological samples with minimal pre-processing, ease of handling and high throughput. Importantly, once the biomarker or combination of biomarkers with potential clinical value has been established, validation analyses can be conducted in close proximity to clinical settings which is important for establishing the utility of new diagnostics in clinical decision making and perhaps future theranostic interventions. This chapter provides protocols for experimental design and methodology aimed at >1) UUdiscovering biologically relevant biomarkers in amniotic fluid using SELDI-TOF; >2) validating the clinical utility of the biomarkers as new diagnostics; >3) translating the biomarker findings into pathophysiological phenomena to provide further insight and extend the current understanding of the disease process. Many of the principles described herein for amniotic fluid could be generalized to studies involving other types of biological samples and other clinical questions.