Silicon microstructures for high-speed and high-sensitivity protein identifications.
Silicon microtechnology has been used to develop a microstructure toolbox in order to enable high accuracy protein identification. During the last 2 years we developed and applied monocrystalline silicon structures and established new automated protein analysis platforms. The development of a high throughput protein platform is presented where fully automated protein identifications are performed. It includes the reduction and alkylation of the protein sample in a standard 96- or 384-well plate format prior to injection of 1 microl samples into the continuous flow based microtechnology platform. The processed sample is transferred to a microchip nanovial array target using piezoelectric microdispensing. Identification is made by MALDI-TOF MS and a database search. After the initial sample reduction and alkylation period of 50 min the platform can digest and process protein samples at a speed of 100 samples in 210 min. An optional configuration of the platform, operating the dispenser in the 'static mode', enables on-target enrichment of low abundant proteins and peptides e.g. from 2DE samples. This makes detection at the low attomole level possible.
Department Electrical Measurement, Lund Institute of Technology, Lund University, Sweden.
T
T
Laurell
J
J
Nilsson
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G
Marko-Varga
D011506:Proteins / Q000737:chemistry*; D012680:Sensitivity and Specificity; D012825:Silicon / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D011506:Proteins; D012825:Silicon