A tumor targeted gene vector modified with G250 monoclonal antibody for gene therapy.
A tumor targeted gene vector modified with G250 monoclonal antibody for gene therapy.
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DOI：
10.1016/j.jconrel.2008.01.016
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发表时间：
2008-04
期刊：
影响因子：
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通讯作者：
Yajun Duan;Junnian Zheng;Sufang Han;Yi Wu;Yanming Wang;Deguan Li;Deling Kong;Yao-ting Yu
中科院分区：
文献类型：
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作者：
Yajun Duan;Junnian Zheng;Sufang Han;Yi Wu;Yanming Wang;Deguan Li;Deling Kong;Yao-ting Yu
G250是一种肿瘤相关抗原，在90%的肾细胞癌（RCC）中发现。为了开发用于RCC基因治疗的高靶向基因载体，制备了G250单克隆抗体，并对其进行了纯化和鉴定。该抗体与聚乙烯亚胺（PEI）化学结合形成IgG-PEI偶联物。该缀合物能够形成纳米大小的DNA复合物，并且具有较窄的尺寸分布。在5种细胞系ket3、Hela、ACHN、HepG2和平滑肌细胞上检测靶向效果和转染效率。采用荧光活化细胞分选（FACS）和荧光素酶测定法定量测定转染量。FACS结果显示，对于G250阳性细胞ket3和Hela， IgG-PEI的转染效率是PEI的2倍。而对于G250阴性细胞，抗体修饰对转染无影响。荧光素酶在ket3细胞中的表达，以酶活性的形式表达，分别比ACHN和SMC高15倍和61倍。在游离抗体存在下，IgG-PEI的靶向作用受损，转染效率归一化。这表明G250抗体是一种理想的靶向配体，可以将基因传递到RCC中。这种IgG-PEI偶联物在RCC基因治疗中的应用将引起人们极大的兴趣。
G250 is a tumor associated antigen that is found on > 90% of renal cell carcinoma (RCC). In order to develop a highly targeting gene vector for RCC gene therapy, G250 monoclonal antibody was prepared, purified and characterized. The antibody was chemically bound to Polyethylenimine (PEI) to form the IgG–PEI conjugate. The conjugate is capable of forming DNA complexes in the size of nano meters and with a narrow size distribution. The targeting effect and transfection efficiency were tested on five cell lines, ketr 3, Hela, ACHN, HepG2, and smooth muscle cells. The transfection was quantitatively determined by fluorescence activated cell sorting (FACS) and luciferase assay. The FACS results show that for G250 positive cells ketr 3 and Hela, the transfection efficiency of IgG–PEI are 2-fold higher than that of PEI. But for G250 negative cells, antibody modification has no effect on transfection. The expression of luciferase in ketr 3 cells which is expressed as enzyme activity is 15-fold and 61-fold higher than that in ACHN and SMC, respectively. In the presence of free antibody, the targeting effect of IgG–PEI is impaired and the transfection efficiency is normalized. It indicates that G250 antibody is an ideal targeting ligand for delivery of genes into RCC. Application of this IgG–PEI conjugate in RCC gene therapy will be of great interest.