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HPLC-MS characterisation of chelate modified somatropin

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HPC-UGent: the central High Performance Computing infrastructure of Ghent University
Abstract
Somatropin is a recombinant human growth hormone, consisting of 191 amino acids. 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 the chelating agent NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid) allows the inclusion of metals coupled to the protein for diagnostic (e.g. 68Ga) or therapeutic (e.g. 90Y) purposes. The NOTA unit is selectively introduced on a lysine side chain. This yields 9 possible labelling sites for somatropin: FPTIPLSRLFDNAMLRAHRLHQLAFDTYQEFEEAYIPKEQKYSFLQNPQTSLCFSESIPTPSNREETQQKSNLELLRISLLLIQSWLEPVQFLRSVFANSLVYGASDSNVYDLLKDLEEGIQTLMGRLEDGSPRTGQIFKQTYSKFDTNSHNDDALLKNYGLLYCFRKDMDKVETFLRIVQCRSVEGSCGF We have applied an enzymatic digestion procedure for the characterisation of the modified somatropin, using trypsin, chymotrypsin and Staphylococcus aureus V-8 proteases. The resulting peptides were then monitored using HPLC-MS2, allowing the characterisation of the modified protein.
Keywords
Somatropin, NOTA, Peptide mapping

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Chicago
Wynendaele, Evelien, Ewald Pauwels, Sylvia Van Dorpe, Valentijn Vergote, Christophe Van De Wiele, and Bart De Spiegeleer. 2010. “HPLC-MS Characterisation of Chelate Modified Somatropin.” In Vlaams Jongerencongres Van De Chemie, 10e, Abstracts. Koninklijke Vlaamse Chemische Vereniging (KVCV).
APA
Wynendaele, E., Pauwels, E., Van Dorpe, S., Vergote, V., Van De Wiele, C., & De Spiegeleer, B. (2010). HPLC-MS characterisation of chelate modified somatropin. Vlaams Jongerencongres van de Chemie, 10e, Abstracts. Presented at the 10e Vlaams Jongerencongres van de Chemie (VJC 10), Koninklijke Vlaamse Chemische Vereniging (KVCV).
Vancouver
1.
Wynendaele E, Pauwels E, Van Dorpe S, Vergote V, Van De Wiele C, De Spiegeleer B. HPLC-MS characterisation of chelate modified somatropin. Vlaams Jongerencongres van de Chemie, 10e, Abstracts. Koninklijke Vlaamse Chemische Vereniging (KVCV); 2010.
MLA
Wynendaele, Evelien, Ewald Pauwels, Sylvia Van Dorpe, et al. “HPLC-MS Characterisation of Chelate Modified Somatropin.” Vlaams Jongerencongres Van De Chemie, 10e, Abstracts. Koninklijke Vlaamse Chemische Vereniging (KVCV), 2010. Print.
@inproceedings{1038333,
  abstract     = {Somatropin is a recombinant human growth hormone, consisting of 191 amino acids. 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 the chelating agent NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid) allows the inclusion of metals coupled to the protein for diagnostic (e.g. 68Ga) or therapeutic (e.g. 90Y) purposes. The NOTA unit is selectively introduced on a lysine side chain. This yields 9 possible labelling sites for somatropin:
FPTIPLSRLFDNAMLRAHRLHQLAFDTYQEFEEAYIPKEQKYSFLQNPQTSLCFSESIPTPSNREETQQKSNLELLRISLLLIQSWLEPVQFLRSVFANSLVYGASDSNVYDLLKDLEEGIQTLMGRLEDGSPRTGQIFKQTYSKFDTNSHNDDALLKNYGLLYCFRKDMDKVETFLRIVQCRSVEGSCGF
We have applied an enzymatic digestion procedure for the characterisation of the modified somatropin, using trypsin, chymotrypsin and Staphylococcus aureus V-8 proteases. The resulting peptides were then monitored using HPLC-MS2, allowing the characterisation of the modified protein.},
  author       = {Wynendaele, Evelien and Pauwels, Ewald and Van Dorpe, Sylvia and Vergote, Valentijn and Van De Wiele, Christophe and De Spiegeleer, Bart},
  booktitle    = {Vlaams Jongerencongres van de Chemie, 10e, Abstracts},
  keyword      = {Somatropin,NOTA,Peptide mapping},
  language     = {eng},
  location     = {Blankenberge},
  publisher    = {Koninklijke Vlaamse Chemische Vereniging (KVCV)},
  title        = {HPLC-MS characterisation of chelate modified somatropin},
  year         = {2010},
}