Development of matrix-assisted ultraviolet laser desorption/ionization mass spectrometry for the structural analysis of glycoproteins

Miscellaneous · Fri Jan 01 00:00:00 EST 1993

OSTI ID:6554224

Chevrier, M R

This thesis describes the design, construction and characterization of an ultraviolet laser desorption time-of-flight [TOF] mass spectrometer and its subsequent application to glycoprotein structural analysis utilizing matrix-assisted laser desorption/ionization [MALDI] mass spectrometry. At the inception of this work, commercial mass spectrometers utilizing MALDI were not available, and most reports of the phenomena utilized the 266 nm wavelength provided by frequency-quadrupled Nd:YAG lasers. This work involved the design and construction of a high-voltage-extraction linear TOF mass analyzer equipped with a multiple sample inlet system and a 337 manometer, 600 picosecond pulsed nitrogen laser. In MALDI the [open quotes]matrix[close quotes], a strong absorber of a laser wavelength, is co-crystallized with the analyte. The laser photons absorbed by the matrix lead to ionization of the analyte and subsequent desorption from the surface into the gas phase. While nicotinic acid and caffeic acid were reported as effective matrices at 266 and 355 nm, respectively, several other matrices were examined for their efficiency at 337 nm, including [alpha]-cyano-4-hydroxy cinnamic acid and gentisic acid, which proved to be advantageous for glycoconjugate analysis. Glycoproteins, phosphoproteins, nucleic acids, and proteolytic digests were all successfully analyzed using the pulsed nitrogen laser. Analysis of numerous peptides and proteins demonstrated femtomolar sensitivity, mass range in excess of 350 kiloDaltons, mass resolution circa 700, and mass accuracy better than 0.1%. The completed instrument was utilized to analyze glycopeptides for carbohydrate sites and microheterogeneity, by performing MALDI mass spectrometry [MALDI/MS] following enzymatic and chemical reactions. In many cases, unfractionated or partially fractionated mixtures were analyzed directly thereby reducing preparative chromatography.

• Research Organization: Johns Hopkins Univ., Baltimore, MD (United States)
• OSTI ID: 6554224
• Resource Relation: Other Information: Thesis (Ph.D.)
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
GLYCOPROTEINS
STRUCTURAL CHEMICAL ANALYSIS
LASER SPECTROSCOPY
FEASIBILITY STUDIES
MASS SPECTROSCOPY
MATRICES
SENSITIVITY
TIME-OF-FLIGHT MASS SPECTROMETERS
DYNAMIC MASS SPECTROMETERS
MASS SPECTROMETERS
MEASURING INSTRUMENTS
ORGANIC COMPOUNDS
PROTEINS
SPECTROMETERS
SPECTROSCOPY
TIME-OF-FLIGHT SPECTROMETERS
550200* - Biochemistry