Écija-Arenas Ángela, Román-Pizarro Vanesa, Fernández-Romero Juan Manuel
Departamento de Química Analítica, Instituto Universitario de Investigación en Química Fina y Nanoquímica (IUNAN), Universidad de Córdoba, Campus de Rabanales, Edificio Anexo "Marie Curie", E-14071, Córdoba, Spain.
Departamento de Química Analítica, Instituto Universitario de Investigación en Química Fina y Nanoquímica (IUNAN), Universidad de Córdoba, Campus de Rabanales, Edificio Anexo "Marie Curie", E-14071, Córdoba, Spain.
Talanta. 2021 Jan 15;222:121532. doi: 10.1016/j.talanta.2020.121532. Epub 2020 Aug 15.
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 (FeO@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.
描述了一种使用多相密度梯度离心（MDGC）结合连续流动系统（CFS）监测混合磁脂质体（h-MLs）分离效率的方法。使用快速溶剂蒸发（RSE）方法合成了几种含有疏水性磁性金纳米颗粒（FeO@AuNPs-C12SH）和不同荧光团的h-MLs悬浮液。MDGC系统是用非线性多相密度梯度制备的，底层为100%（v/v）蔗糖溶液，六层含有蔗糖溶液混合物（浓度范围为10%至55% v/v），以及固定浓度的聚蔗糖（30% v/v）和 Percoll（15% v/v）溶液。密度梯度分布先前使用4480×g的相对离心力（RCF）稳定30分钟。将合成的h-MLs添加到密度梯度分布中，并在2520×g下离心20分钟进行分离。测试分离程序的效率，将分离的提取物吸入CFS中，并在将其转移到引入表面活性剂溶液的检测器之前裂解脂质体。封装的荧光团和其他物质释放提供的发光信号给出了脂质体在每个密度梯度阶段的分布状态。对不同样品的监测显示，每个分离的h-MLs有四个不同的组分（MLs、h-Ls、h-MLs和未封装的荧光团）。还通过共聚焦显微镜获得了关于h-MLs的其他信息。
