Post-translational modifications of recombinant human factor VIII will be determined. Both the """"""""native"""""""" and reduced/aldylated """"""""subunits"""""""" of factor VIII will be digested with tryptin and analyzed by LC/ESIMS and SIM for carbohydrates, phosphate and sulfate esters. The disulfide bridges will be determined from LC/ESIMS data of the unreduced and tryptic digest. The disulfide-containing peptides will be isolated and subjected to MALDI-TOF analysis before and after reduction.
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