Luminescence continuous flow system for monitoring the efficiency of hybrid liposomes separation using multiphase density gradient centrifugation

• Published in: Talanta (Oxford) Vol. 222; p. 121532
• Main Authors: Écija-Arenas, Ángela, Román-Pizarro, Vanesa, Fernández-Romero, Juan Manuel
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15-01-2021
• Subjects: Cell Separation; Centrifugation, Density Gradient; Continuous flow system; Fluorescence; Gold; Liposomes; Liposomes separation; Luminescence; Metal Nanoparticles; Multiphase density gradient centrifugation system; Fluorescence; Liposomes separation; Multiphase density gradient centrifugation system; Continuous flow system
• ISSN: 0039-9140; 1873-3573
• DOI: 10.1016/j.talanta.2020.121532

A method for monitoring the efficiency of the hybrid magnetoliposomes (h-MLs) separation using multiphase density gradient centrifugation (MDGC) coupled with a continuous flow system (CFS) is described. Several h-MLs suspensions containing hydrophobic magnetic gold nanoparticles (Fe3O4@AuNPs-C12SH) and different fluorophores encapsulated have been synthesized using the rapid solvent evaporation (RSE) method. The MDGC system was prepared using a non-linear multiphase density gradient formed with a bottom layer with 100% (v/v) sucrose solution and six layers containing a mixture of sucrose solution (with concentrations ranged between 10 and 55% v/v), and fixed concentrations of ficoll (30% v/v) and percoll (15% v/v) solutions. The density gradient profile was previously stabilized using a relative centrifugal force (RCF) of 4480×g for 30 min. The synthesized h-MLs were added to the density gradient profile and separated by centrifugation at 2520×g for 20 min. The efficiency of the separation procedure was tested, aspirating the separated extract into the CFS and lysing liposomes before their translation to the detector introducing surfactant solutions. The luminescence signals provided by the release of the encapsulated fluorophores and other materials provided the distribution status of the liposomes in each density gradient stage. The monitoring of the different samples revealed four different fractions (MLs, h-Ls, h-MLs, and non-encapsulated fluorophores) for each separated h-MLs. Additional information on the h-MLs has also been acquired by confocal microscopy.
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•A continuous flow system for efficiency evaluation in purification and separation of hybrid magnetoliposomes was developed.•The separation was carried out using a multiphase density gradient centrifugation (MDGC).•The evaluation was performed by coupling the MDGC vial with a continuous flow system equipped with a spectrofluorometer.•Different types of magnetoliposomes containing hydrophobic magnetic NPs and different fluorophores were studied.•The effectiveness of the h-MLs purification was also tested using confocal microscopy images.