APPLICATION OF CAPILLARY ELECTROPHORESIS, HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY, ONLINE ELECTROSPRAY MASS-SPECTROMETRY AND MATRIX-ASSISTEDLASER-DESORPTION IONIZATION-TIME OF FLIGHT MASS-SPECTROMETRY TO THE CHARACTERIZATION OF SINGLE-CHAIN PLASMINOGEN-ACTIVATOR

The analysis of recombinant Desmodus salivary plasminogen activator (D SPA alpha 1), a heterogeneous glycoprotein, is demonstrated through th e use of high-performance liquid chromatography (HPLC), high-performan ce capillary electrophoresis (HPCE), liquid chromatography-electrospra y mass spectrometry (LC-ES-MS), and matrix-assisted laser desorption i onization-time of flight mass spectrometry (MALDI-TOF-MS). The protein is analyzed at three specific levels of detail: the intact protein, p roteolytic digests of the protein, and fractions from the proteolytic digest. A method for ''on-column'' collection of HPLC fractions for su bsequent transfer and analysis by HPCE and MALDI-TOF-MS is shown.