Laser desorption/ionization mass spectrometry MALDI

The most discriminating technique for proving the identity and purity of analyte peak of a chromatogram, especially for analyzing biological samples and natural products, is by using online LC-UV/MS or GC-MS/FTIR methods [15]. Alternatively, one could use a combination of TLC and MS, where direct determination on the TLC plates is made by matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) [16]. 

Many diseases are characterized by the expression of specific proteins1 in some cases, malignant cells yield unique protein profiles when total cellular protein extracts are analyzed by proteomic methods such as two-dimensional gel electrophoresis or matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS).2 High-throughput proteomic studies may be useful to differentiate normal cells from cancer cells, to identify and define the use of biomarkers for specific cancers, and to characterize the clinical course of disease. Proteomics can also be used to isolate and characterize potential drug targets and to evaluate the efficacy of treatments. 

Matrix-assisted laser desorption-ionization mass spectrometry (MALDI-MS), 78 774... 

Lang SR, Staudenmann W, James P, Manz HJ, Kessler R, Galli B, Moser HP, Rummelt A, Merkle HP (1996) Proteolysis of human calcitonin in excised bovine nasal mucosa elucidation of the metabolic pathway by liquid secondary ionization mass spectrometry (LSIMS) and matrix assisted laser desorption ionization mass spectrometry (MALDI). Pharm Res 13 1679-1685. 

Matrix-assisted laser desorption ionization mass spectrometry (MALDI) uses a matrix, typically of aromatic acids, whose molecular weight is similar to those of typical metabolites and thus disallows the mass spectrometric measurement of the latter. 

Mark-Honwink eqnation Relates limiting viscosity number (LVN) to molecular weight LVN = KM . matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) MS in which the sample is placed in a matrix that contains a strong ultraviolet (LTV) absorber chosen to match the UV absorption of the laser, which allows the molecules to become volatilized with minimal fragmentation. 

The exception to this is the application of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). In 1981, Barber and Liu and coworkers independently introduced the concept of employing MALDI where the absorption of the matrix is chosen to coincide with the wavelength of the employed laser to assist in the volatilization of materials. In 1988, Tanaka, Hillenkamp, and coworkers employed the laser as the energy source, giving birth to MALDI-MS. 

Two recently developed mass spectrometric techniques have had a major impact on the analysis of large biomolecules matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and electrospray ionization mass spectrometry (ESI-MS). MALDI-MS was first introduced by Karas and Hillenkamp66 and Tanaka et al.61 in 1988 and has experienced an exponential development. It has become a widespread soft ionization technique for bioorganic samples, especially large biomolecules. Fenn and co-workers68 first published the successful soft ionization technique for... 

Hahner S, Ludemann HC, Kirpekar F, Nordhoff E, Roepstorff P, Galla HJ, Hillenkamp F. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI) of endonuclease digests of RNA. Nucleic Acids Res 1997 25 1957-1964. 

In general, -3% aliquots of 6 of the most symmetrical, latest eluting HPLC absorbance peaks (not also present in the blank digest) were chosen for matrix assisted laser desorption ionization mass spectrometry (MALDI-MS) on... 

Matrix Assisted Laser Desorption Ionization-Mass Spectrometry (MALDI-MS) Mass spectra of native and denatured antibodies were obtained with a PerSeptive Biosystems (Farmingham, MA.) Voyager Elite mass spectrometer operated in the linear mode with a Laser Sciences Inc., 337 nm nitrogen laser. hAB-1 was denatured by boiling the sample in 1.0 M guanidine-HCl, 50 mM Tris pH 7.5 buffer. Native and denatured samples were diluted with 20 mM Tris, 10 mM octylglucoside (Tris/OG) pH 6.8 buffer prior to MALDl-MS analysis. Proteins were spotted on the sample plate as a sandwich between two layers of the matrix. The bottom layer consisted of 100 mM sinapinic acid in acetonitrile and the top layer consisted of 50 mM sinapinic acid in 30% acetonitrile / 70% H2O / 0.07% TFA. The m/z scale of the instrument was calibrated using a Hewlett-Packard protein standard mixture. 

Recombinant wild type hIL-ip, the K138C mutant and the K138C, R4A, L6A triple mutant (mutant 1) were isolated from the soluble fraction of E. coli lysates by ammonium sulfate fractionation and hydrophobic interaction chromatography. The purified proteins were characterized by SDS-PAGE, western blots, N-terminal sequence, size exclusion chromatography (SEC), isoelectric focusing (lEF), matrix assisted laser desorption ionization mass spectrometry (MALDI-MS), and electrospray mass spectrometry (ESMS). 

In this study, we used matrix-assisted laser desorption ionization /Mass Spectrometry (MALDI/MS) to identify the peptides released from gastric parietal cell microsomes. MALDI, because of its sensitivity and relative tolerance to the presence of salts and buffers was examined for the analysis of unfractionated proteolytic digests (9, 10). MALDI with post-source decay (PSD) analysis was used to obtain sequence information on peptides even in crude digestion mixtures. Our strategy (Figure 1) consisted of proteolysis of intact vesicles, centrifugation at high speeds to separate membrane bound and soluble fractions and analysis of the mixture of released peptides by MALDI/MS. In addition, to increase the... 

Also the use of an EWOD system for the automated sample preparation of peptides and proteins for matrix-assisted laser desorption-ionization mass spectrometry (MALDI-MS) was reported. In that work, standard MALDI-MS reagents, analytes, concentrations, and recipes have been demonstrated to be compatible with the EWOD technology, and mass spectra comparable to those collected by conventional methods were obtained [271]. Also a PCR assay has been realized on the platform by temperature cycling of a droplet at rest [272]. Additional informations about the EWOD platform can be found in a comprehensive review [273]. 

SPPS has been enabled through the development of powerful methods for the analysis and subsequent purification of the complex mixture of products typically produced by SPPS. In particular, the development of reversed phase, high performance liquid chromatography (HPLC) and macromolecular mass spectrometry [22], matrix assisted laser desorption/ionization mass spectrometry (MALDI) [23] and electrospray ionization mass spectrometry (ESI-MS)... 

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