DSPE-PEG-NHS介导的蛋白偶联及其对EV交联水凝胶性能的影响-UDP糖丨MOF丨金属有机框架丨聚集诱导发光丨荧光标记推荐西安齐岳生物

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DSPE-PEG-NHS介导的蛋白偶联及其对EV交联水凝胶性能的影响

发布时间：2025-07-08 作者：zyl 分享到：

文献：Extracellular Vesicle Crosslinkers Constructing Hydrogels with Stress-Relaxation and Bioactive Protein Modification

作者：Shi, Yanzhen Jing, Haowen Lu, Fanxuan Zhao, Mengying An, Shuiling Jin, Chang Gao, Yongdong Dai, Yinxin Zhu, Shuxu Yang, Songying Zhang, Xuesong Ye, Xiujun Cai, Yifan Wang

文献链接：https://advanced.onlinelibrary.wiley.com/doi/full/10.1002/admi.202400885

Here, we demonstrate the potential for the sequential insertion of two different DSPE-PEG amphiphilic molecules into the EV membrane: DSPE-PEG-NHS for protein conjugation and DSPE-PEG-SH for hydrogel crosslinking (Figure 4a–c). Using bovine serum albumin (BSA) as a model protein, we show the BSA band shifts upward in the gel band, indicating an increase in molecular weight after conjugation with DSPE-PEG-NHS, verifying the successful preparation of DSPE-PEG-BSA (Figure 4b). We also show that EVs modified with both DSPE-PEG-SH and DSPE-PEG-BSA could be successfully crosslinked with 8-arm PEG-Nb through thiol-ene photopolymerization (Figure 4d). However, the storage modulus of the hydrogels significantly decreased after incorporating DSPE-PEG-BSA, suggesting that the protein may interfere with the EV crosslinking reaction. To further explore protein presentation, we used green fluorescent protein (GFP) as a model protein to visualize its distribution within the hydrogel. Fluorescent imaging revealed that GFP was evenly distributed throughout the EV-crosslinked hydrogels, confirming the successful encapsulation of GFP into the hydrogel network (Figure 4e,f).

我们展示了将两种不同的DSPE-PEG两亲性分子顺序插入EV膜的潜力：DSPE-PEG-NHS用于蛋白质偶联，DSPE-PEG-SH用于水凝胶交联。

使用牛血清白蛋白（BSA）作为模型蛋白，我们显示BSA带在凝胶带中向上移动，表明与DSPE-PEG-NHS结合后分子量增加，验证了DSPE-PEG-BSA的成功制备。

我们还表明，用DSPE-PEG-SH和DSPE-PEG-BSA修饰的EV可以通过硫烯光聚合成功地与8臂PEG-Nb交联。

然而，在掺入DSPE-PEG-BSA后，水凝胶的储能模量显著降低，表明该蛋白质可能会干扰EV交联反应。为了进一步探索蛋白质的呈现，我们使用绿色荧光蛋白（GFP）作为模型蛋白来可视化其在水凝胶中的分布。

荧光成像显示GFP均匀分布在EV交联水凝胶中，证实GFP成功包封到水凝胶网络中。