Liposomes as potential masking agents in sport doping. Part 2: Detection of liposome-entrapped haemoglobin by flow cytofluorimetry
| Autor: | Xavier de la Torre, Monica Mazzarino, Francesco Botrè, Francesco Donati, Sonia Colicchia, Simone Esposito |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Liposome Chromatography 010401 analytical chemistry technology industry and agriculture Pharmaceutical Science Conjugated system 01 natural sciences Fluorescence 0104 chemical sciences Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology chemistry Environmental Chemistry Fluorescein isothiocyanate Haemoglobin-based oxygen carriers Ethylene glycol Spectroscopy Masking agent Whole blood |
| Zdroj: | Drug Testing and Analysis. 9:208-215 |
| ISSN: | 1942-7603 |
| Popis: | 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. Copyright © 2016 John Wiley & Sons, Ltd. |
| Databáze: | OpenAIRE |
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