Silicon microstructures for high-speed and high-sensitivity protein identifications

Silicon microtechnology has been used to develop a microstructure toolbox i n order to enable high accuracy protein identification. During the last 2 y ears we developed and applied monocrystalline silicon structures and establ ished new automated protein analysis platforms, The development of a high t hroughput protein platform is presented where fully automated protein ident ifications are performed. It includes the reduction and alkylation of the p rotein sample in a standard 96- or 384-well plate format prior to injection of 1 mul samples into the continuous flow based microtechnology platform. The processed sample is transferred to a microchip nanovial array target us ing piezoelectric microdispensing. identification is made by MALDI-TOF MS a nd a database search. After the initial sample reduction and alkylation per iod of 50 min the platform can digest and process protein samples at a spee d of 100 samples in 210 min. An optional configuration of the platform, ope rating the dispenser in the 'static mode', enables on-target enrichment of low abundant proteins and peptides e.g. from 2DE samples. This makes detect ion at the low attomole level possible. (C) 2001 Elsevier Science B.V. All rights reserved.