Development of silicon microstructures and thin-film MALDI target plates for automated proteomics sample identifications.
Here we report on the development of a proteomic platform utilizing a piezoelectric flow-through dispensing unit made from silicon microstructures. The use of a novel surface coating, where matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI MS) targets were uniformly precoated with a thin film of matrix/nitrocellulose, made the sample preparation straightforward and enabled the enrichment and analysis of proteins at low levels in proteomics samples. We demonstrate this by analyzing excised spots in a biological sample originating from a human fetal fibroblast cell line that was subjected to 2D gel-electrophoresis. Furthermore, a sample deposition rate below 30 Hz results in an increased analyte density on the dispensed sample spot, rendering signal amplification. In general, the sensitivity for proteins and peptides can be enhanced 10-50 times compared to traditional MALDI sample preparation techniques.
Miliotis
T
T
Department of Analytical Chemistry, Lund University, Box 124, SE-221 00, Lund, Sweden.
Marko-Varga
G
G
Nilsson
J
J
Laurell
T
T
D000818:Animals; D003101:Collodion; D006801:Humans; D009040:Motion Pictures / Q000639:trends*; D011506:Proteins / Q000032:analysis* / Q000235:genetics; D012371:Robotics / Q000295:instrumentation* / Q000379:methods*; D017655:Silicon Compounds; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation* / Q000379:methods* / Q000639:trends
D011506:Proteins; D017655:Silicon Compounds; D003101:Collodion