Liposomes as potential masking agents in sport doping. Part 2: detection of liposome-entrapped haemoglobin by flow cytofluorimetry
Deutscher übersetzter Titel: | Liposomen als potenziell dopingmaskierende Substanzen. Teil 2: Nachweis in Liposomen eingeschlossenen Hämoglobins durch Durchflusszytofluorimetrie |
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Autor: | Esposito, Simone; Colicchia, Sonia; Torre, Xavier de la; Donati, Francesco; Mazzarino, Monica; Botrè, Francesco |
Erschienen in: | Drug testing and analysis |
Veröffentlicht: | 9 (2017), 1/2, S. 208-215, Lit. |
Format: | Literatur (SPOLIT) |
Publikationstyp: | Zeitschriftenartikel |
Medienart: | Elektronische Ressource (online) Gedruckte Ressource |
Sprache: | Englisch |
ISSN: | 1942-7603, 1942-7611 |
DOI: | 10.1002/dta.1956 |
Schlagworte: | |
Online Zugang: | |
Erfassungsnummer: | PU201705003729 |
Quelle: | BISp |
Abstract des Autors
This work presents an analytical procedure for the identification and characterization of liposome-entrapped haemoglobins, based on flow cytofluorimetry. Flow cytofluorimetric detection is carried out following labelling by two distinct fluorescent reagents, an anti-haemoglobin antibody, fluorescein isothiocyanate conjugated, and an anti-poly(ethylene glycol) antibody, streptavidin-phycoerythrin conjugated. This experimental strategy allows the detection of liposome-entrapped haemoglobins in aqueous media, including plasma; the efficacy of the proposed approach has been verified on whole blood samples added with the liposomal formulation (ex-vivo). Additionally, the proposed technique allows the characterization of several key parameters in the study of liposomal haemoglobins, including, for instance (1) the determination of the degree of haemoglobin entrapment by liposomes; (2) the poly(ethylene glycol) insertion efficiency; and (3) the evaluation of liposome-entrapped haemoglobins stability following storage at 4 °C, allowing to follow both the process of haemoglobin loss from liposomes and the liposome degradation. The procedure is proposed for the detection and characterization of liposome-entrapped haemoglobin formulations to control their misuse in sport, but is also suggested for further applications in biological and clinical laboratory investigations.