Disposable polymeric high-density nanovial arrays for matrix assisted laser desorption/ionization-time of flight-mass spectrometry: II. Biological applications.
A novel disposable high-density matrix assisted laser desorption/ionization (MALDI) target plate made either from polymethylmethacrylate (PMMA) or polycarbonate (PC) is presented where thousands (1,200-1,600) of samples can be deposited and subsequently analyzed by MALDI-time of flight (TOF) mass spectrometry. Good reproducibility was obtained across the plate regardless of position on the target plate with a relative standard deviation (RSD) on the peak intensity of typically 30% calculated from data generated by analysis of a 10 nm peptide mixture of angiotensin I, II, III and bradykinin. The nanovial array format combined with microdispensing technology makes it possible to carry out in-vial chemistry on deposited samples. This is demonstrated by the analysis of peptides from beta-casein and subsequent in-vial dephosphorylation of its phosphopeptides at 10 fmol levels by microdispensing of alkaline phosphatase, into the nanovial. The mass spectra obtained from these polymeric targets provides can also be used in high sensitivity applications as shown by peptide mass fingerprinting of human fibroblast proteins separated by two-dimensional gel electrophoresis.
Ekström
S
S
Department of Electrical Measurements, Lund Institute of Technology, Sweden.
Nilsson
J
J
Helldin
G
G
Laurell
T
T
Marko-Varga
G
G
D000595:Amino Acid Sequence; D000809:Angiotensins / Q000032:analysis; D000818:Animals; D001920:Bradykinin / Q000032:analysis*; D002364:Caseins / Q000737:chemistry*; D004209:Disposable Equipment; D015180:Electrophoresis, Gel, Two-Dimensional; D005347:Fibroblasts / Q000737:chemistry*; D006801:Humans; D016547:Kinesins / Q000032:analysis; D008832:Microchemistry / Q000295:instrumentation*; D008840:Microfilament Proteins / Q000032:analysis; D008904:Miniaturization; D025261:Mitochondrial Proton-Translocating ATPases / Q000032:analysis; D008969:Molecular Sequence Data; D036103:Nanotechnology / Q000295:instrumentation*; D010748:Phosphopeptides / Q000032:analysis*; D020543:Proteome; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation*; D014174:Transketolase / Q000032:analysis; D014335:Tropomyosin / Q000032:analysis
D000809:Angiotensins; D002364:Caseins; D008840:Microfilament Proteins; D010748:Phosphopeptides; D020543:Proteome; D014335:Tropomyosin; C071058:moesin; D014174:Transketolase; D025261:Mitochondrial Proton-Translocating ATPases; D016547:Kinesins; D001920:Bradykinin