Dried-droplet probe preparation on AnchorChip targets for navigating the acquisition of matrix-assisted laser desorption/ionization time-of-flight spectra by fluorescence of matrix/analyte crystals.
We have developed a dried-droplet probe preparation method for peptide mass fingerprinting by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS), which uses AnchorChip targets and alpha-cyano-4-hydroxycinnamic acid (CHCA) as a matrix. Upon drying of a matrix and analyte mixture on the AnchorChip, salts and low molecular weight contaminants were pooled at the hydrophilic metal anchor, whereas 10-50 microm matrix/peptide crystals firmly adhered at the surface of a hydrophobic polymer and the entire target could be subsequently washed by submerging it in 5% formic acid for 2-3 min. Epifluorescence microscopy suggested that peptides were completely co-localized with CHCA crystals at the AnchorChip surface. Fluorescent images of the probes were of good contrast and were background-free, compared with images taken by a video camera built into the ion source. CHCA/peptide crystals were easy to recognize at the surface and peptide mass maps were acquired from them without further adjustment of the position of the laser beam. These crystals were remarkably stable towards the laser depletion and almost no matrix-related ions were typically observed in the low m/z region of peptide mass maps. The sensitivity of the peptide mass mapping was at the low-femtomole level.
Thomas
Henrik
H
Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany.
Havlis
Jan
J
Peychl
Jan
J
Shevchenko
Andrej
A
D000818:Animals; D002417:Cattle; D003460:Crystallization; D005453:Fluorescence; D008856:Microscopy, Fluorescence / Q000379:methods; D018715:Microscopy, Video; D010449:Peptide Mapping / Q000295:instrumentation / Q000379:methods*; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation / Q000379:methods*
D012710:Serum Albumin, Bovine