Development of a novel high throughput multiplex affinity capture platform to detect low-abundance cancer related proteins from bodily fluids is proposed. The approach leverages Acoustic Droplet Ejection (ADE) technology from Labcyte Inc., recent developments in biochip fabrication, self-assembled monolayer surface chemistries, and Matrix Assisted Laser Desorption Ionization?Mass Spectrometry (MALDI-MS) and other surface sampling mass spectrometric approaches to create a fully automated, multiplex, affinity capture platform with the reproducibility, sensitivity, and dynamic range required for routine, absolute quantitation of low-abundance cancer-related proteins. In particular, the platform will offer significantly greater throughput than Liquid Chromatography?Mass Spectrometry (LC MS/MS) approaches such as the mass spectrometric quantitation of peptides and proteins using stable isotope standards and capture by anti-peptide antibodies (commonly called the SISCAPA process), while exploiting many of the more attractive aspects of that technique including stable isotope standards and anti-peptide antibodies. With completion of Phase II, the automated platform developed when employed in conjunction with a state-of-the-art MALDI mass spectrometer equipped with a 5 kHz laser will enable multiplex quantitation of protein biomarkers from 96 samples at a rate that is at least 10-fold faster than LC-MS/MS approaches based on Multiple Reaction Monitoring (MRM) and with comparable sensitivity, reproducibility, and dynamic range.
