F. Landry et al., A method for application of samples to matrix-assisted laser desorption ionization time-of-flight targets that enhances peptide detection, ANALYT BIOC, 279(1), 2000, pp. 1-8
Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass
spectrometry has become a fundamental tool for the identification and anal
ysis of peptides and proteins. MALDI-TOF is well suited for the analysis of
complex biological mixtures because samples are crystallized onto a solid
support that can be washed to remove contaminants and salts prior to laser
desorption. A number of approaches for immobilizing samples onto MALDI targ
ets have been put forth. These include the use of different chemical matric
es and the immobilization of samples onto different solid supports. In larg
e part though, the preparation of MALDI targets has been an empirical exerc
ise that often requires a unique series of conditions for every sample. Her
e, a simple method for the application of peptide mixtures onto MALDI targe
ts is put forth. This method differs because peptides are added directly to
a sample of nitrocellulose dissolved in acetone, allowing them to interact
in solution-phase organic solvent. This solution-phase mixture is then spo
tted to the MALDI target and evaporated, forming a homogenous solid surface
for laser desorption. This procedure is robust, highly sensitive, tolerant
to detergents, and easily learned. In our hands, the method provides as mu
ch as a 10-fold enhancement to the detection of tryptic peptide fragments d
erived from in-gel digests. (C) 2000 Academic Press.