Trehalose preserves DDA/TDB liposomes and their adjuvant effect during freeze-drying
Trehalose preserves DDA/TDB liposomes and their adjuvant effect during freeze-drying
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DOI：
10.1016/j.bbamem.2007.05.009
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发表时间：
2007-09-01
影响因子：
3.4
通讯作者：
Agger, Else Marie
中科院分区：
文献类型：
--
作者：
Christensen, Dennis;Foged, Camilla;Agger, Else Marie
二糖是众所周知的在冷冻和干燥期间保护生物结构如蛋白质和基于磷脂的脂质体的试剂。我们研究了两种二糖海藻糖和蔗糖在冷冻干燥时稳定由阳离子二甲基双十八烷基铵（DDA）和海藻糖6，6 '-二山嵛酸酯（TDB）组成的新型非磷脂基脂质体佐剂的能力。使用含有2.5mg/ml DDA和0.5mg/ml TDB以及不同浓度的两种糖的人剂量浓度冷冻干燥脂质体。利用光子相关光谱（PC）研究了再水化对颗粒尺寸的影响，并通过差示扫描量热法（DSC）研究了凝胶到流体的相变。数据显示，浓度高于211 mM的海藻糖在冷冻干燥过程中保护和保存DDA/ TDB，并且脂质体易于再水化。蔗糖作为稳定剂的效率较低，并且必须以高于396 mM的浓度使用以获得相同的效果。用结核病候选疫苗Ag 85 B-ESAT-6与海藻糖稳定的佐剂组合免疫小鼠显示，冻干DDA/TDB脂质体保留了它们刺激强烈的细胞介导的免疫应答和抗体应答的能力。这些发现表明，等渗浓度的海藻糖在冷冻干燥过程中可以保护阳离子DDA/TDB脂质体。由于这是不是基于DDA单独的脂质体的情况下，我们建议，保护是通过直接与头基的TDB和亲液效应的相互作用，而直接与DDA的相互作用起着次要的作用。(C)2007 Elsevier B.V保留所有权利。
Disaccharides are well-known reagents to protect biostructures like proteins and phospholipid-based liposomes during freezing and drying. We have investigated the ability of the two disaccharides trehalose and sucrose to stabilize a novel, non-phospholipid-based liposomal adjuvant composed of the cationic dimethy1dioctadecylammonium (DDA) and trehalose 6,6'-dibehenate (TDB) upon freeze-drying. The liposomes were freeze-dried using a human dose concentration containing 2.5 mg/ml DDA and 0.5 mg/ml TDB with varying concentrations of the two sugars. The influence on particle size upon rehydration was investigated using photon correlation spectroscopy (PCs) and the gel to fluid phase transition was examined by differential scanning calorimetry (DSC). Data revealed that concentrations above 211 mM trehalose protected and preserved DDA/ TDB during freeze-drying, and the liposomes were readily rehydrated. Sucrose was less efficient as a stabilizer and had to be used in concentrations above 396 mM in order to obtain the same effect. Immunization of mice with the tuberculosis vaccine candidate Ag85B-ESAT-6 in combination with the trehalose stabilized adjuvant showed that freeze-dried DDA/TDB liposomes retained their ability to stimulate both a strong cell-mediated immune response and an antibody response. These findings show that trehalose at isotonic concentrations protects cationic DDA/ TDB-liposomes during freeze-drying. Since this is not the case for liposomes based on DDA solely, we suggest that the protection is facilitated via direct interaction with the headgroup of TDB and a kosmotropic effect, whereas direct interaction with DDA plays a minor role. (C) 2007 Elsevier B.V All rights reserved.