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用于毛细管电泳相互作用研究的富含胆固醇的膜涂层:应用于红细胞脂质提取物

Cholesterol-rich membrane coatings for interaction studies in capillary electrophoresis: application to red blood cell lipid extracts.

作者信息

Lindén Maria V, Holopainen Juha M, Laukkanen Antti, Riekkola Marja-Liisa, Wiedmer Susanne K

机构信息

Laboratory of Analytical Chemistry, Department of Chemistry, University of Helsinki, Helsinki, Finland.

出版信息

Electrophoresis. 2006 Oct;27(20):3988-98. doi: 10.1002/elps.200600002.

Abstract

The purpose was to develop a stable biological membrane coating for CE useful for membrane interaction studies. The effect of cholesterol (chol) on the stability of dipalmitoylphosphatidylcholine (DPPC) and sphingomyelin (SM) coatings was studied. In addition, a fused-silica capillary for CE was coated with human red blood cell (RBC) ghost lipids. Liposomes prepared of DPPC/SM with and without chol or RBC ghost lipids were flushed through the capillary and the stability of the coating was measured electrophoretically. Similar mixtures of DPPC/SM with and without chol were further studied by differential scanning calorimetry. The presence of phosphatidylcholine as a basic component in the coating solution of DPPC/SM/chol was found to be essential to achieve a good and stable coating. The results also confirmed the stability of coatings obtained with solutions of DPPC with 0-30 mol% of chol and SM in different ratios, which more closely resemble natural membranes. Finally, the electrophoretic measurements revealed that a stable coating is formed when capillaries are coated with liposomes of RBC ghost lipids.

摘要

目的是开发一种用于毛细管电泳(CE)的稳定生物膜涂层,以用于膜相互作用研究。研究了胆固醇(chol)对二棕榈酰磷脂酰胆碱(DPPC)和鞘磷脂(SM)涂层稳定性的影响。此外,用人类红细胞(RBC)膜脂质对用于CE的熔融石英毛细管进行了涂层处理。将含有或不含chol的DPPC/SM制备的脂质体或RBC膜脂质冲洗通过毛细管,并通过电泳测量涂层的稳定性。通过差示扫描量热法对含有或不含chol的DPPC/SM的类似混合物进行了进一步研究。发现DPPC/SM/chol涂层溶液中磷脂酰胆碱作为基本成分的存在对于获得良好且稳定的涂层至关重要。结果还证实了用含有0 - 30 mol% chol的DPPC溶液和不同比例的SM获得的涂层的稳定性,这些涂层更类似于天然膜。最后,电泳测量表明,当毛细管用RBC膜脂质的脂质体进行涂层处理时会形成稳定的涂层。

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