Rapid identification of proteins by peptide-mass fingerprinting

This review describes the application of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry to carbohydrate analysis and covers the period 1991-1998. The technique is particularly valuable for carbohydrates because it enables underivatised, as well as derivatised compounds to be examined. The various MALDI matrices that have been used for carbohydrate analysis are described, and the use of derivatization for improving mass spectral detection limits is also discussed. Methods for sample preparation and for extracting carbohydrates from biological media prior to mass spectrometric analysis are compared with emphasis on highly sensitive mass spectrometric methods. Quantitative aspects of MALDI are covered with respect to the relationship between signal strength and both mass and compound structure. The value of mass measurements by MALDI to provide a carbohydrate composition is stressed, together with the ability of the technique to provide fragmentation spectra. The use of in-source and post-source decay and collision-induced fragmentation in this context is described with emphasis on ions that provide information on the linkage and branching patterns of carbohydrates. The use of MALDI mass spectrometry, linked with exoglycosidase sequencing, is described for N-linked glycans derived from glycoproteins, and methods for the analysis of O-linked glycans are also covered. The review ends with a description of various applications of the technique to carbohydrates found as constituents of glycoproteins, bacterial glycolipids, sphingolipids, and glycolipid anchors.

Matrix-assisted laser desorption/ionization mass spectrometry of carbohydrates.

Structural investigations of glycoconjugates at high sensitivity.

Molecular Characterization of a Human DNA Kinase

This invention provides methods of retentate chromatography for resolving analytes in a sample. The methods involve adsorbing the analytes to a substrate under a plurality of different selectivity conditions, and detecting the analytes retained on the substrate by desorption spectrometry. The methods are useful in biology and medicine, including clinical diagnostics and drug discovery.

Retentate chromatography and protein chip arrays with applications in biology and medicine

Methods are described fsor the purification of proteins prior to analusis by matrix-assisted laser-desorption mass spectrometry. Contaminated protein sample were immobilized onto the surface of sample targets and rinsed. In general, a layer of electrosprayed nitrocellulose gave better results than the sroughened gold surface of an untreated target. Using this approach, spectra could be obtained from low picomole quantities of protein in the presence of sdcontaminants which did not inhibit the binding of the protein to the substrate.

Sample immobilization protocols for matrix-asssisted laser-desorption mass spectrometry

Publisher Summary This chapter presents a study analyzing sample immobilization protocols for matrix assisted laser desorption mass spectrometry. The mass analyzer used in this work was a Lasermat. The sample targets are formed from stainless steel sheet, and the points irradiated by the laser are confined to a 2 mm diameter central spot. This spot is roughened by bead blasting and flashed with gold so as to provide a wettable surface with many nucleation sites to encourage uniform crystallization of the evaporating sample and matrix solution. The nitrocellulose was applied to the Lasermat sample target by electrospray deposition. A fragment of nitrocellulose membrane was dissolved in acetone to give a concentration of approximately 2 mg/ml. The behavior of the gold and nitrocellulose surfaces was investigated using a range of proteins and contaminants. Reference spectra were obtained from clean material conventionally loaded by mixing sample and matrix solutions on the target. The samples contaminated with 1% sodium azide and 1% SDS gave no useful signals whatsoever when analyzed without purification.

Sample Immobilisation Protocols for Matrix Assisted Laser Desorption Mass Spectrometry

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Sample immobilization protocols for matrix-asssisted laser-desorption mass spectrometry

Sample Immobilisation Protocols for Matrix Assisted Laser Desorption Mass Spectrometry