Effects of employment of distinct strategies to capture antibody on antibody delivery into cultured cells.
Effects of employment of distinct strategies to capture antibody on antibody delivery into cultured cells.
复制标题
采用不同策略捕获抗体对抗体递送至培养细胞的影响。
DOI：
10.1007/s11010-015-2362-x
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发表时间：
2015
期刊：
影响因子：
4.3
通讯作者：
Shinohara Y.
中科院分区：
文献类型：
--
作者：
Kuwahara K;Harada K;Yamagoshi R;Yamamoto T;Shinohara Y.
使用两种递送系统检查抗体递送到培养的HeLa细胞中的特性。这两种系统都使用细胞穿透肽作为抗体侵入细胞的工具，但采用“蛋白A衍生物”或“疏水基序”来捕获抗体。当我们通过使用这两个系统检查Alexa Fluor标记的抗体的摄取时，发现两个系统都有效地将抗体递送到培养的细胞中。然而，当我们将这些系统递送的抗体量与转铁蛋白摄取量进行比较时，前者比后者小10倍。与转铁蛋白摄取相比，抗体递送的效率较低似乎是由于抗体递送试剂的参与，其未能捕获抗体分子。通过使用更大量抗体的实验验证了该解释，并且在该条件下由“蛋白A衍生物”系统递送的抗体量被确定为13 ng蛋白质/105个细胞。通过“蛋白A衍生物”或“疏水基序”实现的抗体递送显示出两个差异，即，递送的抗体分子的细胞内分布的差异和用细胞裂解物观察到的荧光光谱的差异。这两个交付系统之间的这些差异的可能原因进行了讨论。
The characteristics of antibody delivery into cultured HeLa cells were examined using two delivery systems. Both systems used a cell-penetrating peptide as a tool for intrusion of an antibody into the cells, but either a “protein A derivative” or “hydrophobic motif” was employed to capture the antibody. When we examined the uptake of the Alexa Fluor-labeled antibody by the use of these two systems, both systems were found to effectively deliver the antibody into the cultured cells. However, when we compared the amount of antibody delivered by these systems with the amount of transferrin uptake, the former was 10 times smaller than the latter. The lower efficiency of antibody delivery than transferrin uptake seemed to be attributable to the involvement of the antibody delivery reagent, which failed to catch the antibody molecule. This interpretation was validated by an experiment using a larger amount of antibody, and the amount of antibody delivered by the “protein A derivative” system under this condition was determined to be 13 ng proteins/105cells. The antibody delivery achieved by the “protein A derivative” or “hydrophobic motif” showed two differences, i.e., a difference in intracellular distribution of the delivered antibody molecules and a difference in the fluorescence spectrum observed with cellular lysates. Possible reasons for these differences between the two delivery systems are discussed.