ATTACHMENT OF ANTIBODIES TO STERICALLY STABILIZED LIPOSOMES - EVALUATION, COMPARISON AND OPTIMIZATION OF COUPLING PROCEDURES
ATTACHMENT OF ANTIBODIES TO STERICALLY STABILIZED LIPOSOMES - EVALUATION, COMPARISON AND OPTIMIZATION OF COUPLING PROCEDURES
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DOI：
10.1016/0005-2736(95)00138-s
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发表时间：
1995-11-01
影响因子：
3.4
通讯作者：
ALLEN, TM
中科院分区：
文献类型：
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作者：
HANSEN, CB;KAO, GY;ALLEN, TM
先前已经开发了几种用于将抗体（Ab）结合至脂质体的偶联方法。我们感兴趣的是，如果这些方法中的一些将是适合于连接抗体的脂质体（SL）的长循环配方，空间稳定的聚（乙二醇）（PEG），我们研究了三个“经典”的耦合方法，其中抗体连接在双层表面的SL，和两个新的耦合方法，其中抗体连接在PEG末端。检查的参数包括结合效率、抗体表面密度、免疫脂质体远程装载抗癌药物阿霉素的能力以及所得免疫脂质体与靶细胞的特异性结合。非共价生物素-抗生物素蛋白偶联方法导致细胞表面的低Ab密度，其中马来酰亚胺衍生的Ab通过掺入脂质体中的硫醇化磷脂附着于脂质体表面的偶联方法也是如此。在这些方法中获得的低水平Ab可能是由于PEG干扰Ab进入脂质体表面。然而，当将马来酰亚胺衍生化的Ab偶联到硫醇化的PEG时，将偶联反应远离脂质体表面，实现了非常高的偶联效率，并且这些免疫脂质体实现了与其靶细胞的良好特异性结合。氧化Ab的Fc区并通过腙键将其偶联至PEG末端是效率较低的偶联方法，但具有保留Ab取向的优点。有效的远程加载的阿霉素被发现的免疫脂质体，其中抗体连接在PEG末端。
Several coupling methods for binding antibodies (Ab) to liposomes have previously been developed. We were interested in examining if some of these methods would be suitable for attaching Ab to long-circulating formulations of liposomes (SL), sterically stabilized-with poly(ethylene glycol) (PEG), We studied three 'classical' coupling methods in which Ab was attached at the bilayer surface of sL, and two new coupling methods in which Ab was attached at the PEG terminus. Parameters examined included binding efficiency, antibody surface density, the ability of the immunoliposomes to remote-load the anticancer drug doxorubicin, and the specific binding of the resulting immunoliposomes to target cells. The non-covalent biotin-avidin coupling method resulted in low Ab densities at the cell surface, as did a coupling method in which maleimide-derivatized Ab was attached to the liposome surface through a thiolated phospholipid incorporated into the liposomes. The low levels of Ab achieved in these method was likely due to interference by PEG with the access of the Ab to the liposome surface. However, when a maleimide-derivatized Ab was coupled to thiolated PEG, moving the coupling reaction away from the liposome surface, very high coupling efficiencies were achieved, and these immunoliposomes achieved good specific binding to their target cells. Oxidizing the Fc region of the Ab and coupling it to the PEG terminus through a hydrazone bond was a less efficient coupling method, but had the advantage of retaining Ab orientation. Efficient remote-loading of doxorubicin was found for immunoliposomes in which Ab was attached at the PEG terminus.