Calicheamicin derivatives (MW approximately 1500) and monoclonal antibodies (MoAbs) conjugated to calicheamicin derivatives (MW approximately 150,000) were analyzed by UV-MALDI/MS, IR-MALDI/MS, and ESI/MS. These materials are potent anticancer agents. Calicheamicin derivatives and conjugates rapidly degrade upon UV irradiation but are relatively stable during IR irradiation and under ESI conditions. A unique feature of IR-MALDI/MS is a 2 times enhancement in resolution relative to UV-MALDI/MS for masses above approximately 50,000 Da resulting in a molecular ion envelope containing a series of partially resolved peaks of the calicheamicin-MoAb conjugates. The mass shift difference between the peak maxima corresponded to the mass change due to the covalent addition of calicheamicin derivatives to the monoclonal antibody. The distribution of the calicheamicin derivatives in the monoclonal antibodies was computed by deconvoluting the partially resolved peak envelope. A unique feature of the ESI mass spectra, under unit resolution conditions, is that the distribution of the carbohydrates can be well resolved for pure MoAbs and can be only partially resolved for conjugated MoAbs. Average loading values for calicheamicia derivatives when conjugated to MoAbs were computed from UV-MALDI/MS, IR-MALDI/MS, and ESI/MS data and the results compared with the average loading values obtained by UV absorption spectrometry. Very low average loading values were computed from UV-MALDI/MS data due to the degradation of the conjugated calicheamicin derivatives during the UV irradiation process. The IR-MALDI/MS average loading values, obtained with glycerol as the matrix, were consistent with the UV absorption spectrometry values for conjugates having hydrolytically stable linkers, but not when the linker contained a hydrolytically labile hydrazone. ESI/MS average loading values were generally lower than the corresponding values obtained by IR-MALDI/MS. The average loading values and distributions obtained using IR-MALDI/MS were more reliable than the corresponding ESI/MS values because the partially resolved, singly and doubly charged peaks in the IR-MALDI spectra can be mathematically deconvoluted, while the overlapping, highly multiply charged peaks of the electrospray spectra can only be partially deconvoluted.
Two series of SnO thin films, one doped with N and one doped with H, were deposited on c-plane sapphire by reactive ion beam sputter deposition starting from growth parameters optimized for stoichiometric SnO. The amounts of dopants incorporated into the SnO:H and SnO:N samples were quantified by secondary ion mass spectroscopy. The influence on the structural and electrical properties of SnO thin films was studied as a function of dopant concentration. In the case of N doping, all N incorporated, probably as NO, are active as the acceptor and exhibit long-term stability. We assign an acceptor activation energy of 100 to 150 meV to NO. However, we observe a change in the film morphology at a critical N concentration of about 7⋅1017cm−3, which deteriorates the structural properties of the films. In the case of SnO:H, the situation is different. We observe an outdiffusion of H after growth, i.e., the samples are not stable in the long term. Nevertheless, all H incorporated up to a H-content of 1019cm−3 seem to be electrically active and exhibit an activation energy between 150 and 250 meV, likely corresponding to Hi. Furthermore, at H contents above 1019cm−3, we observe molecular H2 inside the SnO:H thin films. We conclude that N doping of SnO is better suited for tuning the p-type conductivity of SnO. However, it will be essential to overcome the morphology change observed at the critical N concentration to fully explore the tunability of the p-type conductivity in device applications.
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