Characterization of tyrosine sulfate residues in antihemophilic recombinant factor VIII by liquid chromatography electrospray ionization tandem mass spectrometry and amino acid analysis

Rapid Commun Mass Spectrom. 1999;13(11):1016-23. doi: 10.1002/(SICI)1097-0231(19990615)13:11<1016::AID-RCM599>3.0.CO;2-5.


Recombinant Factor VIII (rFVIII) is involved in the cascade of biochemical reactions leading to blood coagulation and is used for the treatment of haemophilia A. Plasma-derived FVIII (pdFVIII) has been reported to be post-translationally modified by sulfation of tyrosine residues at positions 346, 1664, 1680, 718, 719 and 723. This report describes the quantitation of tyrosine sulfate residues in BHK-derived, human rFVIII by amino acid composition analysis and the identification of their positions in the polypeptide sequence using a combination of liquid chromatography and electrospray ionization mass spectrometry in the analysis of proteolytic digests of the protein.

MeSH terms

  • Amino Acid Sequence
  • Amino Acids / analysis
  • Chromatography, High Pressure Liquid
  • Factor VIII / analysis*
  • Hydrolysis
  • Mass Spectrometry
  • Molecular Sequence Data
  • Peptide Fragments / analysis
  • Peptide Mapping
  • Recombinant Proteins / analysis
  • Spectrophotometry, Ultraviolet
  • Thrombin / chemistry
  • Tyrosine / analogs & derivatives*
  • Tyrosine / analysis


  • Amino Acids
  • Peptide Fragments
  • Recombinant Proteins
  • tyrosine O-sulfate
  • Tyrosine
  • Factor VIII
  • Thrombin