Avidin-biotin immobilization of unilamellar liposomes in gel beads for chromatographic analysis of drug-membrane partitioning

被引:76
作者
Yang, Q
Liu, XY
Ajiki, S
Hara, M
Lundahl, P
Miyake, J
机构
[1] Agcy Ind Sci & Technol, Natl Inst Adv Interdisciplinary Res, Bion Design Grp, Tsukuba, Ibaraki 305, Japan
[2] Stanley Elect Co Ltd, Tsukuba Res Lab, Tsukuba, Ibaraki 30026, Japan
[3] Agcy Ind Sci & Technol, Natl Inst Biosci & Human Technol, Tsukuba, Ibaraki 305, Japan
[4] Uppsala Univ, Ctr Biomed, Dept Biochem, S-75123 Uppsala, Sweden
来源
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 1998年 / 707卷 / 1-2期
关键词
avidin-biotin immobilization; unilamellar liposomes; drug-membrane partitioning;
D O I
10.1016/S0378-4347(97)00620-8
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
To construct a homogeneous lipid membrane chromatographic phase, biotinylated unilamellar liposomes of small and large sizes (SUVs and LUVs, respectively) were immobilized in avidin-or streptavidin-derived gel beads in amounts up to 55 mu mol phospholipid/ml gel bed at yields above 50%. The immobilized liposomes exhibited excellent stability due to avidin-biotin multiple-site binding. The trapped volume and size distribution of the immobilized liposomes (0.33-0.42 mu l/mu mol lipid and 20-30 nm diameter for SUVs, 1.7-1.9 mu l/mu mol lipid and 80-120 nm for LUVs) indicated the unilamellarity and integrity of the immobilized liposomes. Partitioning of 15 pharmaceutical drugs into the bilayers of LUVs immobilized in different eel matrices correlated very well, as shown by chromatographic drug retention analysis. The partitioning of several beta-blockers into the immobilized LUVs showed a close correlation with their partitioning, reported in the literature, into free liposomes. The avidin-biotin-immobilized unilamellar liposomes can thus be used for chromatographic analysis and screening of solute-membrane interactions. (C) 1998 Elsevier Science B.V.
引用
收藏
页码:131 / 141
页数:11
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