3D Printed Integrated Sample Preparation and Chromatography Platform for Limited Volume Proteomics

Abstract Limited volume (microliters down sub nanoliters) complex biological sample processing is routine in the growing fields of proteomics. The complexity of the proteome, the dynamic concentration range of proteins, and the absence of PCR-like amplification techniques make sample processing a crucial cog in this field. However, the limitations of existing sample processing technologies, including low throughput, high cost, and sub-optimal processing, affect data quality and have checked its widespread use in biology and medicine. This project will employ custom 3D printable carbon microbead stationary phase materials and additive manufacturing tools to fabricate integrated solid-phase biological processing platforms. Its overall goal will be to prepare microbead solid phase arrays to extract, fractionate, and separate proteins from single-cell or limited-volume lysate samples. The proposed technology and products will be scalable, low-cost, reusable, require minimal reagents, and allow high throughput efficient protein pre-fractionation and separation. Additional features will allow facile interfacing with upstream lysate preparation systems and sensors downstream to significantly improve proteomics methods' speed, sensitivity, and dynamic range. This project's successful outcomes will dramatically advance the development of bespoke integrated single-cell and small- volume complex biological sample processing platforms. At scale, such capabilities would realize small- volume, single-cell proteomics, or single-molecule protein sequencing. These biotools will rapidly accelerate biomarker discovery for the early detection of diseases and infections.