Advanced search
1 file | 222.69 KB Add to list

Characterisation of modified pharmaceutical proteins: the somatropin case

Evelien Wynendaele (UGent) , Daphne Wiskerke (UGent) and Bart De Spiegeleer (UGent)
Author
Organization
Abstract
Somatropin, a recombinant protein containing 191 amino acids, is derived from the endogenous human growth hormone, somatotropin [1]. This protein is clinically used in children and adults with inadequate endogenous growth hormone to stimulate a normal bone and muscle growth. In addition, somatropin is currently being investigated for the diagnosis and radiotherapy of certain hormonal cancers. The modification of the protein with a chelating agent like NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid) allows the inclusion of metals coupled to the protein. The NOTA unit is selectively introduced on a lysine side chain. This yields 9 possible labelling sites for somatropin: 38, 41, 70, 115, 140, 145, 158, 168 and 172. As site-specific labelling is necessary to avoid active region interactions, characterisation of the chelate-modified somatropin is indispensable. Therefore, we have applied an enzymatic digestion procedure using trypsin, chymotrypsin and Staphylococcus aureus V-8 proteases. The resulting peptides were then monitored using HPLC-MSn, allowing the investigation of the exact position of amino acid modifications.
Keywords
NOTA, Somatropin, Site-specific labelling

Downloads

  • 2010-265c Poster scientific afternoon FFW.pdf
    • full text
    • |
    • open access
    • |
    • PDF
    • |
    • 222.69 KB

Citation

Please use this url to cite or link to this publication:

MLA
Wynendaele, Evelien, Daphne Wiskerke, and Bart De Spiegeleer. “Characterisation of Modified Pharmaceutical Proteins: The Somatropin Case.” Scientific Afternoon, Posters. 2010. Print.
APA
Wynendaele, E., Wiskerke, D., & De Spiegeleer, B. (2010). Characterisation of modified pharmaceutical proteins: the somatropin case. Scientific Afternoon, Posters. Presented at the Scientific Afternoon 2010.
Chicago author-date
Wynendaele, Evelien, Daphne Wiskerke, and Bart De Spiegeleer. 2010. “Characterisation of Modified Pharmaceutical Proteins: The Somatropin Case.” In Scientific Afternoon, Posters.
Chicago author-date (all authors)
Wynendaele, Evelien, Daphne Wiskerke, and Bart De Spiegeleer. 2010. “Characterisation of Modified Pharmaceutical Proteins: The Somatropin Case.” In Scientific Afternoon, Posters.
Vancouver
1.
Wynendaele E, Wiskerke D, De Spiegeleer B. Characterisation of modified pharmaceutical proteins: the somatropin case. Scientific Afternoon, Posters. 2010.
IEEE
[1]
E. Wynendaele, D. Wiskerke, and B. De Spiegeleer, “Characterisation of modified pharmaceutical proteins: the somatropin case,” in Scientific Afternoon, Posters, Ghent, Belgium, 2010.
@inproceedings{1038335,
  abstract     = {Somatropin, a recombinant protein containing 191 amino acids, is derived from the endogenous human growth hormone, somatotropin [1]. This protein is clinically used in children and adults with inadequate endogenous growth hormone to stimulate a normal bone and muscle growth. In addition, somatropin is currently being investigated for the diagnosis and radiotherapy of certain hormonal cancers. The modification of the protein with a chelating agent like NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid) allows the inclusion of metals coupled to the protein. The NOTA unit is selectively introduced on a lysine side chain. This yields 9 possible labelling sites for somatropin: 38, 41, 70, 115, 140, 145, 158, 168 and 172. As site-specific labelling is necessary to avoid active region interactions, characterisation of the chelate-modified somatropin is indispensable. Therefore, we have applied an enzymatic digestion procedure using trypsin, chymotrypsin and Staphylococcus aureus V-8 proteases. The resulting peptides were then monitored using HPLC-MSn, allowing the investigation of the exact position of amino acid modifications.},
  author       = {Wynendaele, Evelien and Wiskerke, Daphne and De Spiegeleer, Bart},
  booktitle    = {Scientific Afternoon, Posters},
  keywords     = {NOTA,Somatropin,Site-specific labelling},
  language     = {eng},
  location     = {Ghent, Belgium},
  title        = {Characterisation of modified pharmaceutical proteins: the somatropin case},
  year         = {2010},
}