Identification of endothelial proteins by MALDI-MS using a compact disc microfluidic system.
Vascular endothelial proteins have been analyzed using two-dimensional (2D) gel electrophoresis and subsequent mass spectrometry, with separate methods for the intervening sample preparations. Compact disc (CD) technology was found to be rapid, giving high overall yield both with ordinary Coomassie staining and with Sypro Ruby staining. Combined with automatic in-gel digestion, the CD technology has great capacity for large numbers of protein analysis, although for limited sample numbers, manual methods can give similar sequence coverage. In a test set of 48 samples, 45 proteins were identified using the CD preparation technique, 32 identified with higher sequence coverage using the CD technique, 7 with higher using ZipTips in a robotic workstation, and 5 with higher coverage using dried droplets of unpurified samples. In the process of these methodological comparisons, basic patterns for 116 endothelial proteins were defined, representing 297 separate protein spots on the 2D gels.
Hirschberg
Daniel
D
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, SE-171 77 Stockholm, Sweden.
Tryggvason
Sam
S
Gustafsson
Magnus
M
Bergman
Tomas
T
Swedenborg
Jesper
J
Hedin
Ulf
U
Jörnvall
Hans
H
D016250:Compact Disks; D015180:Electrophoresis, Gel, Two-Dimensional; D042783:Endothelial Cells / Q000737:chemistry; D004730:Endothelium, Vascular / Q000737:chemistry*; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D011506:Proteins