POROUS LAYER OPEN TUBULAR COLUMNS WITH IMMOBILIZED TRYPSIN FOR PROTEIN DIGESTION

Summary We have developed a monolithicporous layer open tubular (PLOT) column with immobilized trypsin for protein digestion. The PLOT column was prepared in a 10 Pm ID fused silica capillary. Trypsin was immobilized on the monolithicsurface and the developed enzyme reactor was used for protein digestionfollowed by on-line ESI/MS analysis. 1. Introduction Immobilized microfluidic enzyme reactors (IMER) have shown a high potential in a wide variety of research fields such as medical diagnostics, organic synthesis, drug discovery, or biosensors 1,2 . The use of immobilized enzymes, especially of proteases, has many advantages, e.g., very short digestion time, elimination of autodigestion and therefore absence of undesirable protease fragments in the resulting peptide mixture. The reactors with immobilized enzymescan also be integrated online into the multidimensional systemsenabling automated high-throughput protein analysis. A variety of methods are now available for immobilization of enzymes on the solid supports such as particles or monolithiccolumns