NDLI: Quantification of pharmaceutical peptides using selenium as an elemental detection label

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Metallomics Vol - 6 Issue - 8

Metallomics Vol - 6 Issue - 7

Metallomics Vol - 6 Issue - 6

Metallomics Vol - 6 Issue - 5

Metallomics Vol - 6 Issue - 4

Metallomics Vol - 6 Issue - 3

Metallomics Vol - 6 Issue - 2

Quantification of pharmaceutical peptides using selenium as an elemental detection label

Content Provider	Royal Society of Chemistry (RSC)
Author	Bahnsen, Jesper Søborg Gammelgaard, Bente Stürup, Stefan Franzyk, Henrik Gabel-Jensen, Charlotte Møller, Laura Hyrup
Copyright Year	2014
Abstract	The aim of the present work was to demonstrate how selenium labelling of a synthetic cell-penetrating peptide may be employed in evaluation of stability and quantitative estimation of cellular uptake by inductively coupled plasma mass spectrometry (ICP-MS). Two analogues of the cell-penetrating peptide, penetratin, were synthesized, one with selenomethionine (SeMet) added at the N-terminal of the peptide (N-PenMSe) and the other with the internal methionine (Met) replaced with SeMet (i-PenMSe). The purity of the synthesized peptides was 92% for N-PenMSe and 89% for i-PenMSe as determined by liquid chromatography (LC)-ICP-MS. The selenium-labelled peptides were investigated by cell uptake studies in HeLa WT cells. The stability of the peptides was monitored in water, cell medium and during cell uptake studies. Total uptake of selenium was quantified by flow injection (FI)-ICP-MS. Speciation analysis of cell samples by LC-ICP-MS showed mainly uptake of the intact peptides, while the amount of intact peptides in cell lysates was semi-quantitatively determined. The selenium-containing penetratin analogues were to some extent degraded in pure cell medium, while an extensive degradation was observed during cell uptake studies. The major degradation products were determined by LC-electrospray ionization mass spectrometry (ESI-MS). The labelling method in combination with FI-ICP-MS, LC-ICP-MS and LC-ESI-MS techniques provided detailed information on the fate of penetratin in cellular uptake studies. Most pharmaceutical peptides, including penetratin, are synthetic analogues of endogenous peptides, and incorporation of selenium may improve the critical assessment of the native drug or drug delivery candidate early in the drug development process.
Starting Page	1639
Ending Page	1647
Page Count	9
File Format	HTM / HTML PDF
ISSN	17565901
Volume Number	6
Issue Number	9
Journal	Metallomics
DOI	10.1039/c4mt00085d
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Methionine Drug delivery High-performance liquid chromatography HeLa N-terminus Ionization Selenium Inductively coupled plasma mass spectrometry Peptide Mass spectrometry Speciation Cell-penetrating peptide Selenomethionine
Content Type	Text
Resource Type	Article
Subject	Chemistry Medicine Metals and Alloys Biochemistry Biomaterials Biophysics

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