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  • Title: Mapping of recombinant hemoglobin using immobilized trypsin cartridges.
    Author: Lippincott J, Hess E, Apostol I.
    Journal: Anal Biochem; 1997 Oct 15; 252(2):314-25. PubMed ID: 9344419.
    Abstract:
    A tryptic mapping procedure has been developed for a recombinant hemoglobin (rHb1.1) using an immobilized trypsin cartridge. Apohemoglobin is passed through the trypsin cartridge and the products of the digestion are captured directly onto an in-line C18 reversed-phase column. The peptides are then separated using a gradient elution. This new procedure is rapid and reproducible and can be fully automated. The total time of analysis is less than 2 h. The mapping of apohemoglobins produced an unexpected isomerization of two peptides: beta8,9 (K66-K82) and alpha8,9 (K61-K90). It appears that the isomerization may occur through transpeptidation followed by proteolysis at a newly generated site next to the site of ligation. This mapping procedure can be a useful tool for research and routine analysis of proteins.
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