Mass-selected peptide ions produced by electrospray ionization were deposited onto fluorinated self-assembled monolayer surfaces (FSAM) surfaces by soft landing using a Fourier transform ion cyclotron resonance mass spectrometer (FT-ICR MS) specially designed for studying interactions of large ions with surfaces. Analysis of the modified surface was performed in situ by combining 2-keV Cs+ secondary ion mass spectrometry with FT-ICR detection of the sputtered ions (FT-ICR-SIMS). Regardless of the initial charge state of the precursor ion, the SIMS mass spectra included singly protonated peptide ion, peptide fragment ions, and peaks characteristic of the surface in all cases. In some experiments, multiply protonated peptide ions and [M + Au]+ ions were also observed upon SIMS analysis of modified surfaces. For comparison with the in situ analysis of the modified surfaces, ex situ analysis of some of the modified surfaces was performed by 25-keV Ga+ time-of-flight-secondary ion mass spectrometry (TOF-SIMS). The ex situ analysis demonstrated that a significant number of soft-landed peptide ions remain charged on the surface even when exposed to air for several hours after deposition. Charge retention of soft-landed ions dramatically increases the ion yields obtained during SIMS analysis and enables very sensitive detection of deposited material at less than 1% of monolayer coverage. Accumulation of charged species on the surface undergoes saturation due to coulomb repulsion between charges at close to 30% coverage. We estimated that close to 1 ng of peptide could be deposited on the spot area of 4 mm2 of the FSAM surface without reaching saturation.

Protein ions, after mass spectrometric separation, can be soft-landed into liquid surfaces with preservation of their native structures. Retention of biological activity is strongly favored in glycerol-based surfaces but not in self-assembled monolayer solid surfaces. Soft-landing efficiency for multiply-charged hexokinase ions was found to be some four times higher for a glycerol/fructose liquid surface than for a fluorinated self-assembled monolayer surface. Soft-landing into liquid surfaces is also shown to allow (1) protein purification, (2) on-surface identification of the soft-landed material using MALDI, and (3) protein identification by in-surface tryptic digestion. Pure lysozyme was successfully isolated from different mixtures including an oxidized, partially decomposed batch of the protein and a partial tryptic digest. Liquid glycerol/carbohydrate mixtures could be used directly to record MALDI spectra on the soft-landed compounds provided they were fortified in advance with traditional MALDI matrices such as p-nitroaniline and ␣-cyano-4-hydroxycinnamic acid. Various proteins were soft-landed and detected on-target using these types of liquid surface. Soft-landing of multiply-charged lysozyme ions onto fluorinated self-assembled monolayer surfaces was found to occur with a limited amount of neutralization, and trapped multiply-charged ions could be desorbed from the surface by laser desorption. Initial data is shown for a new approach to protein identification that combines top-down and bottom-up approaches by utilizing protein ion soft-landing from a protein mixture, followed by tryptic digestion of the landed material and detection of characteristic tryptic fragments by MALDI. , has been applied in experiments ranging from studies using chemically inert and structurally organized self-assembled monolayers (SAMs) as substrates for ion storage [2], to studies of ion mobility through ice and vapor deposited films [3,4], to experiments aimed at developing a separation method in a form of preparatory mass spectrometry [5,6]. Simple organic cations [7,8] [12][13][14] have all been the targets of examination by ion soft-landing. The separation, purification, and storage of chemical species by a completely non-destructive, mass spectrometric approach are the objectives of the studies currently underway in our laboratory.In a preliminary report [5], we described a method which showed the potential for generating biologically active protein chips from protein mixtures by massselective ion soft-landing. After ionization of the mixture by electrospray (ESI), individual protein ions were mass-selected and soft-landed at predetermined locations on the surface. The results showed that the use of mass spectrometry as a separation method in biology provides high selectivity because components with different molecular formulas can be separated and softlanded. However, retention of the biological activity of a protein or other biomolecule in the course of this (or any mass spectrometry experiment) is a much more chall...

Ion/surface collision phenomena in the hyperthermal collision energy regime (1-100 eV) are reviewed, with emphasis on chemical processes associated with the impact of small organic and biological ions at functionalized self-assembled monolayer surfaces. Inelastic collisions can lead to excitation of the projectile ion and can result in fragmentation, a process known as surface-induced dissociation which is useful in chemical analysis using tandem mass spectrometry. Changes in charge can accompany ion/surface collisions and those associated with a change in polarity (positive to negative ions or vice versa) are an attractive method for ion structural characterization and isomer differentiation. The surface-induced charge inversion of nitrobenzene and other substituted aromatics is discussed. Reactive collisions occurring between gaseous ions and surfaces depend on the chemical nature of the collision partners. These reactions can be used for selected chemical modifications of surfaces as well as for surface analysis. Particular emphasis is given here to ion soft-landing, another type of ion/surface interaction, in which the projectile ion is landed intact at the surface, either as the corresponding neutral molecule or, interestingly but less commonly, in the form of the ion itself. The ion soft-landing experiment allows for preparative mass spectrometry; for example the preparation of pure biological compounds by using the mass spectrometer as a separation device. After separation, the mass-selected ions are collected by soft-landing, at different spatial points in an array. If the experiment is performed using a suitable liquid medium, in the case of some proteins at least, biological activity is retained.

Mass-selected peptide ions produced by electrospray ionization were deposited as ions by soft-landing (SL) onto fluorinated and hydrogenated self-assembled monolayer (FSAM and HSAM) surfaces using a Fourier transform ion cyclotron resonance mass spectrometer (FT-ICR MS) specially designed for studying collisions of large ions with surfaces. Analysis of modified surfaces was performed in situ by combining 2 keV Cs(+) secondary ion mass spectrometry with FT-ICR detection of the sputtered ions (FT-ICR-SIMS). Similar SIMS spectra obtained following SL at different collision energies indicate that peptide fragmentation occurred in the analysis step (SIMS) rather than during ion deposition. The effect of the surface on SL was studied by comparing the efficiencies of SL on gold, FSAM, HSAM, and COOH-terminated SAM surfaces. It was found that FSAM surfaces are more efficient in retaining ions than their HSAM analogues, consistent with their larger polarizability. The efficiency of soft-landing of different peptides on the FSAM surface increases with the charge state of the ion, also consistent with an ion-polarizable molecule model for the initial stage of soft-landing on SAM surfaces. The gradual decrease of peptide ion deposition efficiency with an increase in collision energy found experimentally was quantitatively rationalized using the hard-cube model.

No abstract