APPLYING CHARGE DISCRIMINATION WITH ELECTROSPRAY-IONIZATION MASS-SPECTROMETRY TO PROTEIN ANALYSES

Electrospray ionization with a magnetic sector mass spectrometer and scanning array detector has unique advantages for sensitive analyses of large biomolecules. The ability tb discriminate against low charge state ions (smaller peptides, buffers and salts, background ions) allows for detection of more highly charged ions from proteins present at much lower concentration relative to the small ions from buffers and detergents present. Low femtomole detection limits can be achieved for proteins greater than 100 ku. The charge discrimination phenomenon is more pronounced for higher charged ions, and especially for large biomolecules. Although the charge distribution for the monomer (66 ku) and dimer (133 ku) species of bovine serum albumin overlap, both species can be ascertained readily in a mixture because the lower charged monomer ions have higher optimum microchannel plate voltages than the higher charged dimer ions. Protein-containing solutions can be analyzed directly by electrospray ionization-mass spectrometry (ESI-MS) with array detection, which eliminates time-consuming separation and sample cleanup procedures. For example, heme-containing proteins can be directly detected from ESI-MS of human blood (hemoglobin) as well as from raw meat juices (hemoglobin and myoglobin).