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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).

Gso6k8KkkM7RycDaQNRv.png

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。

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