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Disposable polymeric high-density nanovial arrays for matrix assisted laser desorption/ionization-time of flight-mass spectrometry: II. Biological applications

Författare:
Publiceringsår: 2001
Språk: Engelska
Sidor: 3984-3992
Publikation/Tidskrift/Serie: Electrophoresis
Volym: 22
Nummer: 18
Dokumenttyp: Artikel
Förlag: Wiley - VCH Verlag GmbH

Sammanfattning

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 (1200-1600) 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.

Disputation

Nyckelord

  • Chemistry
  • polymer high-density matrix assisted laser desorption
  • ionization target
  • plates
  • phosphoproteins
  • peptide mapping
  • array
  • microdispensing
  • IMMOBILIZED PH GRADIENTS
  • PROTEIN IDENTIFICATION
  • SAMPLE PREPARATION
  • 2-DIMENSIONAL ELECTROPHORESIS
  • LIQUID-CHROMATOGRAPHY
  • SEQUENCE
  • DATABASES
  • PURIFICATION
  • PROTEOMICS
  • EXPRESSION
  • MEMBRANE

Övriga

Published
Yes
  • ISSN: 0173-0835

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