Matrix-analyte-interaction in MALDI-MS: Pellet and nano-electrospray preparations

Horneffer V, Gluckmann M, Kruger B, Karas M, Strupat K, Hillenkamp F

Abstract

The incorporation of analytes into matrix crystals and even more so its mechanistic aspects as a prerequisite for a successful MALDI-MS has been discussed controversially in the literature. Solventless sample preparation techniques can shed new light on this question. In order to investigate some crucial aspects of these preparation techniques, lyophylized peptides and proteins were ground or milled with the powder of two different matrices, 2,5-DHB as incorporating matrix and 2,6-DHB for which protein incorporation was definitely excluded in a prior study, and pressed into pellets. The dependence of the quality of the UV-MALDI-spectra on the mass (up to 12,360 Da) and the milling time in a ball mill is reported. For mellitin different initial axial ion velocities were found, when desorbed from 2,5-DHB-pellets as prepared and after wetting and re-drying. Velocities of 150 and 580 m s(-1) for dry and wetted pellets are taken as representative for hard desorption from a surface and soft desorption of matrix-incorporated analytes, respectively. Proteins labeled with either fluorescein isothiocyanate (FITC) or Texas Red (TR) were nano-electrosprayed onto a bed of ferulic acid in a 'dry' or 'wet' mode. All 'dry' deposits exhibit strong fluorescence but do not yield MALDI-ion signals. All 'wet' deposits yield MALDI-signals of the proteins; the fluorescence of FITC is quenched in 'wet' deposits because of the low matrix pH. (c) 2005 Elsevier B.V. All rights reserved.