文献：Exosomal Vaccine Loading T Cell Epitope Peptides of SARS-CoV-2 Induces Robust CD8+ T Cell Response in HLA-A Transgenic Mice

作者：An-Ran Shen 1, Xiao-Xiao Jin 2, Tao-Tao Tang 1, Yan Ding 2, Xiao-Tao Liu 2, Xin Zhong 1, Yan-Dan Wu 2, Xue-Lian Han 3, Guang-Yu Zhao 3, Chuan-Lai Shen 2, Lin-Li Lv 1,✉, Bi-Cheng Liu 1

文献链接：https://pmc.ncbi.nlm.nih.gov/articles/PMC9346304/

摘要：

We used the same procedure used for generating the BSA-PEG-lipid-exosome complex to generate the peptide-PEG-lipid-exosome complex. DMPE-PEG-NHS and the mixture of VEPs were dissolved in DMSO at a 1:3 molar ratio with 1% TEA overnight at RT to form the peptides-PEG-DMPE complex. Then, HEPES buffer was added to dilute the DMSO concentration below 1%. Then, DSPE-PEG was added at a 1:1 molar ratio to the pre-existing peptides-DMPE-PEG-NHS complex and maintained for 15 min at 60℃. The free peptides were removed, and the protein levels in the resultant peptides-PEG-micelles were quantified using the BCA assay kit and stored at 4℃ for use in the following 2 weeks. Then, the peptides-PEG-micelles were redissolved, ultrasonicated to reduce the micelle size, and co-incubated with the RBC-derived exosomes at a 1:1 ratio in a water bath for 2 h at 40℃. The resulting solution was cooled to 4℃ followed by instant purification by the FPLC system with the SXK16/40 Smartarose CL-4B filtration column. Finally, the fractions of the second peak were collected, concentrated in normal saline by ultrafiltration, and used as the peptides-PEG-lipid-exosome complex.

我们使用与产生BSA-PEG脂质外泌体复合物相同的程序来产生肽-PEG-脂质外泌物复合物。

将DMPE-PEG-NHS和VEP的混合物以1:3的摩尔比与1%TEA在室温下溶解在DMSO中过夜，形成肽-PEG-DMPE复合物。

然后，加入HEPES缓冲液以将DMSO浓度稀释至1%以下。然后，将DSPE-PEG以1:1的摩尔比加入到预先存在的肽DMPE-PEG-NHS复合物中，并在60℃下保持15分钟。

去除游离肽，使用BCA检测试剂盒定量所得肽-PEG胶束中的蛋白质水平，并将其储存在4℃下，以便在接下来的2周内使用。

然后，将肽-PEG胶束重新溶解，超声处理以减小胶束尺寸，并在40℃的水浴中以1:1的比例与RBC衍生的外泌体共孵育2小时。

将所得溶液冷却至4℃，然后用SXK16/40 Smartrose CL-4B过滤柱通过FPLC系统立即纯化。最后，收集第二个峰的组分，通过超滤在生理盐水中浓缩，用作肽-PEG-脂质外泌体复合物。

相关推荐：

DSPE-Chitosan

DSPE-RB

DSPE-FITC

DSPE-ICG

DSPE-OPSS

DSPE-PCL

DSPE-PEI

DSPE-PEI-Dextran

DSPE-PLA

DSPE-PLGA

DSPE-Poly(L-glutamic acid)

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