EFFECTS OF TRITON-X-100 CONCENTRATION AND INCUBATION-TEMPERATURE ON CARBOXYFLUORESCEIN RELEASE FROM MULTILAMELLAR LIPOSOMES
EFFECTS OF TRITON-X-100 CONCENTRATION AND INCUBATION-TEMPERATURE ON CARBOXYFLUORESCEIN RELEASE FROM MULTILAMELLAR LIPOSOMES
复制标题
DOI：
10.1016/0005-2736(86)90211-7
复制
发表时间：
1986-07-24
期刊：
影响因子：
--
通讯作者：
WEINER, N
中科院分区：
文献类型：
--
作者：
SILA, M;AU, S;WEINER, N
羧基荧光素是测量囊泡内容物释放速率的最常用的探针。这种方法获得的数据的有效性取决于用去污剂(通常是Triton X-100)处理脂质体时，基于染料完全释放的终点的获得。然而，Triton并不完全从多层脂质体中释放包裹的羧基荧光素，洗涤剂处理时标志物的释放量和释放速度是脂质体的脂组成、Triton浓度、温度和洗涤剂孵育时间的函数。Distearoyl-L-.alpha.-phosphatidylcholine/cholesterol(2：1，摩尔%)多层脂质体经0.5%Triton在22度处理后的荧光“终点”。C(一种经常使用的条件)只有在72度下用5%Triton处理的脂质体的五分之一左右。处理条件似乎影响从部分或完全破坏的脂质体的脂类中释放羧基荧光素，以及随后将游离染料分配到水相中。这种效应会导致对多层脂质体稳定性数据的解释出现严重错误。然而。Triton允许在所有测试条件下从小的单层囊泡中完全释放包裹的染料。
Carboxyfluorescein is the most commonly used probe to measure the rate of release of vesicle contents. The validity of the data obtained by this method depends on obtaining an end point based on the complete release of the dye on treatment of the liposomes with a detergent, usually Triton X-100. However, Triton does not completely release entrapped carboxyfluorescein from multilamellar liposomes and the amount and rate of release of marker upon detergent treatment is a function of lipid composition of the liposome, Triton concentration and temperature and duration of detergent incubation. The fluorescence ''end point'' for distearoyl-L-.alpha.-phosphatidylcholine/cholesterol (2:1, mol%) multilamellar liposomes treated with 0.5% Triton at 22.degree. C (a condition often used) is only about one-fifth the value for liposomes treated with 5% Triton at 72.degree. C. The conditions of treatment appear to affect the release of carboxyfluorescein from the lipid of the partially or completely disrupted liposome and the subsequent partitioning of the free dye into the aqueous phase. This effect can lead to serious errors in the interpretation of multilamellar liposome stability data. However. Triton allows complete release of entrapped dye from small unilamellar vesicles under all conditions tested.